摘要
为进一步揭示蓖麻毒素的毒理作用机制,本研究从蓖麻毒素的基本生物学特征(等电点、分子量、氨基酸序列、细胞毒性)分析入手,对其毒性作用的发挥及相关作用机理进行深入探讨。以小鼠为研究对象,在口服蓖麻毒素72h以后,病变反应最为明显,采用HE染色病理切片、透射电镜、扫描电镜观察等技术对蓖麻毒素的肠道及免疫器官损伤作用进行详尽分析;鉴于镜下观察的细胞凋亡状况,本研究从凋亡的基本特征入手(线粒体膜电位、形态学特征、DNA“ladder”等),对蓖麻毒素可诱导免疫细胞凋亡的机制进行研究。首先检测凋亡细胞胞内活性氧(ROS)水平,继而分析了与ROS有密切级联反应的丝裂原活化蛋白激酶(MAPKs)三条通路的激活反应,胞外调节激酶(ERK1/2)、应激激活蛋白激酶(P38)、c-jun氨基末端激酶(JNK1/2 ),并进一步做阻断分析。结果显示,蓖麻毒素诱导淋巴细胞的凋亡是通过激活P38/MAPK来实现的。鉴于蓖麻毒素对小鼠肠道的强损伤作用,本研究利用蛋白质组学研究手段(2-DE、SPR、HPLC、HPCE-ESI-MS)在体外对小肠刷状缘膜蛋白(BBMP)与蓖麻毒素之间的作用以及互作强度做具体分析,筛选到28种与蓖麻毒素具有互作效应的蛋白。
Ricin (RT) is the most toxic natural poisons of plant proteins up to now. One molecule ricin enters into cell can kill it as a consequence of protein synthesis inhibition. The toxicity of ricin relies on its unique structure, which contains two chains, RTA and RTB. RTA is the active chain with N-glycoside activity, it can cleave the N-glycosidic bond of a single adenine residue in a defined position of 28S rRNA (A4324) within 60S large subunit in the ribosome of eukaryotic cell, and hydrolyzes N-glycosidic bond of A4324 to remove adenine. 28s rRNA will be degraded because of the loss of RNase resistance and can not combine with EF-2, thereby deterring the formation of ribosome, EF-2, GTP complex to lead to the inhibition of protein synthesis. RTB with lectin activity is able to identify and combine the end of the receptors containing galactose structure on the cell surface to assist the RTA to enter into the cell. The present study found that ricin toxicity on cells is not only manifested as inhibition of protein synthesis, but also induced apoptosis of immune cells and played an extremely significant role in intestinal injury. In order to further reveal the toxicity mechanism of ricin, our subject start from the basic biological characteristics of ricin and discuss in-depth on the toxic effects and its related mechanism.
Firstly, isoelectric point (PI) and molecular weight (MW) of purified ricin were analyzed by the capillary electrophoresis. It indicated that the PI and MW were 6.49 and 61 900, respectively. Due to the different places of production and growing conditions possibly, they differed slightly from the former reported results 7.1 and 64,000. Secondly, detected amino acid sequence of ricin, and 24 aa mutated compared with the sequence of former reported. Thirdly, detected the toxicity of ricin in mice, median lethal dose (LD50) was 5.0mg/kg by oral administration and 4.6μg/kg by intraperitoneal injection. At last, detected its cytotoxicity, lymphacytes were treated with different concentrations (0, 0.3, 3, 30 and 300μg/ml) of ricin and different handling durations (3, 6, 12 and 24h), which indicated that the cells showed proliferative response obviously at the first 3 hours and the viability was higher than the control groups, but the viability began to decrease after infecting 4 hours. At 0.3μg/ml of ricin when treated with higher concentration of ricin, the viability decreased obviously without the progress of proliferative response and the reduction extent was enhancing along with the lengthen of handling time. That showed the cytotoxic effect had concentration-depended and tine-depended. While, the IC50 of lymphacytes was 3μg/ml.
The most obvious pathological changes were found in mice after oral administrated 72 hours. They showed depression, anorexia, lethargy shock and coma in clinical, with swollen stomach, obvious congestive reaction in duodenum, hydroabdomen, and intestinal lesions. HE staining showed that necrosis and falling of the intestinal epithelial cells, mesocaval fibrin leaked out the duodenum and jejunum, mucosa intrinsic layer and the central lymphatic lacteal expanded, there was the existence of thrombosis on the lamina propria and the grass-roots level, but there were no lesions in ileum. In addition, atrophy of spleen and thymus was apparent. Under the transmission electron microscope, apoptosis debris of lymphocyte could be seen clearly, in addition, the color of splenic corpuscle in spleen changed light, and some parts of the splenic corpuscle were intumescent, lymphocyte proliferated, multinucleated giant cells (hematopoietic stem cells) came out, and the thymus bleed. In the lesioned immune cells, heterochromatin showed the phenomena of margination, edge were thicker, the membrane structure came out inside of the nuclear, the chromatin around the nuclear presented crescent-shaped. Duodenal intestinal almost completely lost by observing with scanning electron endoscopic, it indicated the ricin had a strong role in intestinal injury.
In the second experiment we could see the apoptotic immune cells, but do not make a decisive judgment to determine that this apoptosis was caused by ricin. So in this experiment, we began with the basic characteristics of apoptosis to further study the mechanism of apoptosis induced by ricin. It did dot contradict with the tradition theory that ricin as a protein synthesis inhibitor, and most likely, it might be the crucial complement to the existing apoptotic theories. Firstly, we used flow cytometry to detect mitochondrial membrane potential with DePsipherTM as a detection probe. Results showed 3μg/ml ricin treated cells for 3 h, and 0.3% H2O2 treated cells for 5 min as positive control. Generally, change of MMP determined by FCM couold be expressed by the ratio of red fluorescence intensity to green (Fig.3). In the present study, ratio of treated cells (2.37) was significantly lower than control cells (3.31) (p<0.5). Ratio deflection illustrated the beginning of the apoptosis. Mitochondrial membrane potential cells were an important indicator of apoptosis, once the membrane potential change, the entire apoptotic process was irreversible. At the 3μg/ml ricin treated cells for 6h and after Hoechst33258 staining, the treated cells shrinkage, chromatin condensation, nuclear brokedown and produced apoptotic bodies, membrane became blurred, incomplete, etc. In the DNA "ladder" detection, ricin-treated cells (3μg/ml ricin for 0h, 24h and 36h) showed typical internucleosomal DNA fragmentation or‘‘ladder’’formation at the two time points tested , which indicated that cell genomic DNA degradation, and the intensity of banding was more prominent at 36h, control cells showed a clear band without DNA fragmentation. Several reports had suggested that high ROS concentrations significantly inhibited cell growth and induced apoptosis, so we observed the ROS level in ricin-treated lymphocytes. 3μg/ml ricin treated cells for 15-120 min, and 0.3% H2O2 treated cells for 5min as positive control. After treated for 15min, ROS level increased about 3-fold compared with control groups. After 30min, it reached the maximum level, and no further increase at longer treating durations. Results showed that ROS level had intimated relationship with the treating durations. There was evidence indicated that ROS regulated metabolic processes by acting as intracellular messenger molecules and that ROS mediate specific signaling pathways by modulating gene expression through regulation of several transcription factors and activation of mitogen-activated protein kinases (MAPKs), in view of the cascade relationship between them, we exploited Western-blot to study the signal transpathway about apoptosis. MAPKs contained three major transpathways, extracellular-regulated kinase (ERK1/2), stress-activated protein kinase (P38), c-jun N-terminal kinase (JNK1/2). In order to identify the process of apoptosis was conducted by which transpathway, we did the blocking analysis by using their specific inhibitors (SP600125, SB-202190, U0126), and final result showed that ricin-induced lymphocyte apoptosis was possibly by activating P38/MAPKs.
For the severe damage of ricin to small intestinal, we developed a series of research to further study this intoxic effect. On passive and/or active transport, most toxins or drugs developed for oral administration were absorbed in the gastrointestinal tract followed by systemic circulation. Because in vivo investigations were, time-consuming and often difficult to quantify the results, it was evident that the predicative in vitro modle was needed. In a present study, we described a novel method to estimate binding events occurring on intestinal membranes based on the technical platform of proteomics (2-DE, SPR, HPLC, HPCE-ESI-MS). After the extraction and purification of mouse intestinal brush border membrane proteins, we did the kinetics analysis based on SPR technology, it could rapidly offer valuable information on the rate and extent of adsorption, association/dissociation kinetics, and the affinity constants of specific ligand-receptor interactions. Results showed that the affinity kinetics (KA) was 1.5×107 , disassociation kinetics(KD)was 6.64×10-8 . This result provided a strong theoretical basis for the explanation of intestinal injury leaded by ricin. In the following test, we analyzed the complexity of brush border membrane protein according to 2-DE technology. Though it had been purified, its composition was still very complicated, so in order to avoid blind identification of a single point, we used SPR recovery unit to recover the interacted components with real-time detection by HPLC. Then, the mixed composition was identified by LC-MS-MS. Results showed that 28 proteins were interacted with ricin in brush border membrane, including the KDEL endoplasmic reticulum retention signal receptor, annexin, original upstream regulatory binding protein, vacuolar protein, etc. This result not only provide a new evidence and a research window for the mechanism of intestinal injury induced by ricin, also set up a new platform to clarify the specific toxic effect of ricin.
引文
[1]Lord J M, Roberts L M, Robertus J D, et al. Ricin: strcture, mode of actin and some current applications. FASEB J, 1994, 8: 201~209.
[2]Robert B, John F H, Mark A, et al. Ricin: mechanism of action, detection and intoxication. J Toxiol, 1995, 14: 483~491.
[3]Franz D R, Aaz N K. Medical aspects of chemical and biologicalwarfare, chapter 32, ricin toxin, fall church. VA: Office of the Surgeon General, 1997: 631~638
[4]Stillmark. Ueber Ricen. Arbeiten des Pharmacologischen Institutes zu Dorpat, iii, 1889. Cited in: Flexner. The histological changes produced by ricin and abrin intoxications. J Exp Med. 1897;2:197~216.
[5]Cawley D B, Hedbolm M L, Hoffman E J, et al. Differential sensitivity of rat liver and wheat germ ribosomes to inhibition of polyuridylic acid translation by ricin A chain. Arch Biochem Biophys, 1977; 182: 690~695
[6]Ehrlich P. Experimentelle Untersuchungenüber Immunit?t, I: Euber Ricin. Deutsch Med Wochenschr. 1891;17:976~979
[7]Olsnes S, Pihl A. Abrin, ricin, and their associated agglutinins. In: Cuatrecasas P, ed. Receptors and Recognition: The Specificity and Action of Animal, Bacterial, and Plant Toxins. Chapman and Hall; 1976: 129~173.
[8]Ghosh, P. C., Wu, H. C., 1988. Enhancement of cytotoxicity of modeccin by nigericin in modeccin-resistant mutant cell lines. Exp. Cell. Res. 174, 397-410.
[9]Magerstadt M. Therapeutic aminoconjugates. In: Antibody Conjugates and Malignant Disease. Boca Raton, Fla: CRC Press; 1991: Chap 3.
[10]Ucken F, Frankel A. The current status of immunotoxins: An overview of experimental and clinical studies as presented at the 3rd International Symposium on Immunotoxins. Leukemia. 1993;7:341~348
[11]Vitetta E, Thorpe P, Uhr J. Immunotoxins: Magic bullets or misguided missiles? Trends Pharmacol Sci. 1993;14(5):148~154.
[12]Vitetta E, Krolick K, Muneo M, Cushley W, Uhr J. Immunotoxins: A new approach to cancer therapy. Science. 1983;219:644~649.
[13]Thorpe PE, Mason DW, Brown AN, et al. Selective killing of malignant cells in leukemic rat bone marrow using an antibody–ricin conjugate. Nature . 1982;297:594~596.
[14]Cookson J, Nottingham J. A Survey of Chemical and Biological Warfare. New York, NY: Monthly Review Press. 1969: 6.
[15]Ramesh Hegde. Studies of the variants of the protein toxins Ricin and Abrin. Eur. J.Biochem.1992, 204, 155~164.
[16]Bernhijmer A W. Perspectives in toxinology, Abrin and Ricin: two toxic tectins inactivating eukaryotic ribosome. 1977, sju R. Olsnes(eds), Awiley Medical publication.
[17]Bharadwaj, S., Rathore, S. S., Ghosh, P. C., 2006. Enhancement of the cytotoxicity of liposomal ricin by the carboxylic ionophore monensin and the lysosomotropic amine NH4Cl in Chinese hamster ovary cells. Int. J. Toxicol. 25, 349-59.
[18]Robertus J. Toxin structure. In: Frankel A, ed. Immunotoxins. Boston, Mass: Kluwer Academic Publishers; 1988: 11~24.
[19]Dietrich J B, Ribereau G G.Identity of the N-terminal sequences of the three A chains of mistletoe lectins: homology with ricin-like plant toxin and single-chain ribosome inhibiting proteins. Anticancer Drugs, 1992, 3: 507~511.
[20]Dimitriadis, G. J., Butters, T. D., 1979. Liposome-mediated ricin toxicity in ricin-resistant cells. FEBS. Lett. 98, 33-6.
[21]Katzin B J, Collins E J, Robertus J D. Structure of ricin A-chain at 2.5 A°. Proteins, 1991; 10: 251~259.
[22]Mlsna D, Monzingo A F, Katzin B J, et al. Structure of recombinant ricin A-chain at 2.3 A°resolution. Protein Sci 1993; 2: 429~435.
[23]Weston SA, Tucker AD, Thatcher DR, et al. X-ray structure of recombinant ricin A-chain at 1.8 A°resolution. J Mol Biol 1994; 244: 410~22.
[24]Burlet A, Chapleur C M, Haumont P B. Long-term reduction of vasopressin excretion induced by the central injection of an immunoconjugate. Neruoscience, 1992, 50: 965~973.
[25]Endo Y, Tsurugi K. RNA N-glycosidase activity of ricin A-chain: mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. J Biol Chem 1987; 262: 8128~8130.
[26]Lewis M S, Youle R J. Ricin subunit association: thermodynamics and the role of the disulfide bond. J Biol Chem, 1986; 261: 11571~11577.
[27]Wright H T, Robertus J D. The intersubunit disulfide bridge of ricin is essential for cytotoxicity. Arch Biochem Biophys,1987; 256: 280~284.
[28]Endo Y, Tsurugi K. The RNA N-glycosidase activity of ricin A-chain. J Biol Chem 1988; 263: 8735~8739.
[29]Morris K N. Proc. Natl. Acad. Sci. USA, 1992,89:48~69.
[30]Ban N, Nissen P, et al. Placement of protein and RNA structures into a 5A°resolution map of the 50S ribosomal subunit. Nature 1999; 400: 841~847.
[31]Sperti S, Montanaro L, Mattioli A, et al. Inhibition by ricin of protein synthesis in vitro: 60 S ribosomal subunit as the target of the toxin. Biochem J, 1973; 136: 813~815.
[32]Nilsson L, Nygard O. The mechanism of the protein synthesis elongation cycle in eukaryotes: effect of ricin on the ribosome interaction with elongation factors. Eur J Biochem, 1986; 161: 111~117.
[33]Hartley M R, Legname G, Osborn R, et al. Single-chain ribosome-inactivating proteins from plants depurinate Escherichia coli 23S ribosomal RNA. FEBS Lett 1991; 290: 65~68.
[34]Villafranca J E, Robertus J D. Ricin B chain is the product of gene duplication. J Biol Chem, 1981; 256: 554~556.
[35]Ramakrishnan S F, Mohavraj D. Cytotoxic conjugates containing translational inhibitory proteins, Ann Rev Pharmacol Toxicol, 1992, 32: 579.
[36]Magnusson S., Kjeken R.Characterization of two distinct pathways of endocytosisof ricin by rat liver endothelial cells. Exp.Cell.Res.,1993,205: 118~125.
[37]Riccobono F., Fiani M. L. Mannose receptor dependent uptake of ricin A1 and A2 chains by macrophage. Carbohydr Res,1996, 282(2): 285~292.
[38]Barbieri L, Valbonesi P, Bonora E, et al. Polynucleotide: adenosine glycosidase activity of ribosome-inactivating proteins: effect on DNA, RNA and poly A. Nucleic Acids Res, 1997; 25: 518~522.
[39]Brigotti M, Alfies R, Sestili P, et al. Damage to nuclear DNA induced by Shiga toxin 1 and ricin in human endothelial cells. FASEB J, 2002; 16: 365~372.
[40] T. Nakanishi, Y. Nishi, E.F. Sato, M. Ishii, T. Hamada, M. Inoue, Thermal injury induces thymocyte apoptosis in the rat, J.Trauma 44 (1998) 143~148.
[41]van Damme E, Hao Q, Chen Y, et al. Ribosome-inactivating proteins: a family of plant proteins that do more than inactivate ribosomes. Crit Rev Plant Sci, 2001; 20: 395~465
[42]Barbieri L, Valbonesi P, Bonora E. Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins: effect on DNA, RNA and poly A. Nucleic Acids Res, 1997; 25: 519~522
[43]Peumans W J, Hao Q, van Damme E J. Ribosome-inactivating proteins from plants: more that RNA N-glycosidases? FASEB J, 2001; 15: 1493~1506
[44]Balint G A. Ricin: The toxic protein of castor oil seeds. Toxicology, 1974;2(1): 77~102.
[45]Fu T. ,Burbage C. and Tagge E. ,Double - lectin site ricin B chain mutants espressed in insect cells have residual galactose binding : evidence for more than two lectin site on the ricin toxin B chain.Bioconjug Chem, 1996 ;7 (6) :651~658.
[46]张今,鲁小青,陈百先,等.蓖麻蛋白糖脂脂体对肝癌细胞的杀伤作用及其理化特殊性.上海铁道大学学报,1998,19 (9):290—291.
[47]Wannemaker R. Assistant Division Chief, Toxinology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Md. Personal communication, September 1994.
[48]Eiklid K, Olsnes S, Pihl A. Entry of lethal doses of abrin, ricin and modeccin intothe cytosol of HeLa cells. Exp Cell Res 1980;126:321~326.
[49]Butterworth A G, Lord J M. Ricin and Ricinus communis agglutinin subunits are all derived from a single-size polypeptide precursor. Eur J Biochem 1983; 137: 56~62.
[50]Ferrini J B, Martin M, Taupiac M P, et al. Expression of functional ricin B chain using the baculovirus system. Eur J Biochem, 1995; 233: 772~777.
[51]Lamb F I, Roberts LM, Lord J M. Nucleotide sequence of cloned cDNA coding for preproricin. Eur J Biochem, 1985; 148: 265~270.
[52]Lord J M. Precursors of ricin and Ricinus communis agglutinin: glycosylation and processing during synthesis and intracellular transport. Eur J Biochem 1985; 146: 411~416.
[53]Roberts L M, Lord J M. The synthesis of Ricinus communis agglutinin. Eur J Biochem, 1981; 119: 31~34.
[54]Lord J M. Synthesis and intracellular transport of lectin and storage protein precur sors in endosperm from castor bean. Eur J Biochem 1985; 146: 403~409.
[55]Lord J M, Harley S M. Ricinus communis agglutinin B chain contains a fucosylated oligosaccharide side chain not present on ricin B chain. FEBS Lett, 1985: 189; 72~76
[56]Harley S M, Lord J M. In vitro endoproteolytic cleavage of castor bean lectin precursors. Plant Sci, 1985; 41: 111~116.
[57]Frigerio L, Vitale A, Lord J M, et al. Free ricin A chain, proricin and native toxin have different cellular fates when expressed in tobacco protoplasts. J Biol Chem, 1998; 273: 14194~14199.
[58]Frigerio L, Jolliffe N A, Di Cola A, et al. The internal propeptide of the ricin precursor carries a sequence-specific determinant for vacuolar sorting. Plant Physiol, 2001; 126: 167~175.
[59]Lambert J M. , Goldmacher V S. ,and Collinson A R. ,et al. An Immuntoxin prepared with blocked ricin :a natural plant toxin adaped for therapeutic use. Cancer Res. 1991 ;51 (23) :6236~6242
[60]Sharma DK et al. Selective stimulation of caveolar endocytosis by glycosphingolipids and cholesterol. Mol Biol Cell 2004; 15:3114~3122.
[61]Jolliffe N A, Ceriotti A, et al. The position of the proricin vacuolar targeting signal is functionally important. Plant Mol Biol, 2003; 41: 825~835.
[62]Ramesh Hegde, et al. Studies of the variants of the protein toxins Ricin and Abrin. Eur. J.Biochem.1992, 204, 155~164.
[63]Sjur Olsnes, et al. Different biological properties of the two constituent peptide chains of ricin, a toxic protein inhibiting protein synthesis. Biochemistry, 1973,12: 3121~3126.
[64]Jon D, et al. Crystallization of Ricin A chain obtained from a cloned gene expressed in Escherichia coli. Biol Chem.,1987,262: 19~20.
[65]Hare O M, et al. Expression of Ricin A chain in Escherichia coli. FEBS Lett., 1987,216: 73~78.
[66]Hailling K, et al. Genomic cloning and characterization of a ricin gene from Ricinus communis Nucl.. Acids Res., 1985,13: 8019~8033.
[67]Tagge E P, et al. Preproricin expressed in Nicotiana tobacum cells in vitro is fully processed and biologically active. Protein Exprpurif, 1996,8: 109~118.
[68]Richardson P T, Westby M, et al. Recombinant proricin binds galactose but does not depurinate 28S ribosomal RNA. FEBS Lett 1989; 255: 15~20.
[69]Lewis M S, Youle R J. Ricin subunit association: thermodynamics and the role of the disulfide bond. J Biol Chem, 1986; 261: 11571~11577.
[70]Wright H T, Robertus J D. The intersubunit disulfide bridge of ricin is essential for cytotoxicity. Arch Biochem Biophys, 1987; 256: 280~284.
[71] Fagan S M, Beevers H. Ricin inhibition of in vivo protein synthesis by plant ribosomes. Proc Natl Acad Sci, 1982; 79: 5935~5938.
[72]Taylor S, Massiah A, Lomonossoff G, et al. Correlation between the activities of five ribosome-inactivating proteins in depurination of tobacco ribosomes and inhibition of tobacco mosaic virus infection. Plant J, 1994; 5: 827~835.
[73]Ghosh P, Wellner R, Cragoe E, Wu H. Enhacement of ricin cytotoxicity inChinese hamster ovary cells by depletion of intracellular K+: Evidence for Na+/H+ exchange system in Chinese hamster ovary cells. J Cell Biol, 1985,101: 350~357.
[74]Moya M, Dautry-Varsat A, Goud B, Louvard D, Boquet P. Inhibition of coated pit formation in Hep2 cells blocks the cytotoxicity of diptheria toxin but not that of ricin toxin. J Cell Biol, 1985, 101(2): 548~559.
[75]Sandvig K, Olsnes S, Pihl A. Kinetics of binding of the toxic lectins abrin and ricin to surface receptors of human cells. J Biol Chem, 1976, 251(13): 3977~3984.
[76]Audi J, Belson M, Patel M, Schier J, Osterloh J. Ricin poisoning: a 659 comprehensive review. JAMA 2005;294(18):2342~51.
[77]Hudson T, Neville D. Temporal separation of protein toxin translocation from processing events. J Biol Chem, 1987, 262:16484~16494.
[78]Crompton R. Georgi Markov: Death in a pellet. Med Leg J, 1980, 48: 51~62.
[79]Olsnes S, Pihl A. Toxic lectins and related proteins. In: Cohen P, van Heyningen S, eds. Molecular Action of Toxins and Viruses. Amsterdam, Netherlands: Elsevier Biomedical Press; 1982: 51~105.
[80]Wilhelmsen C, Pitt L. Lesions of acute inhaled lethal ricin intoxication in rhesus monkeys. Vet Pathol, 1993, 30: 482
[81]Moisenovich,AlexT,NataliaM, et al. Endosomal ricin transport :involvement of Rab4- and Rab5-positive compartments. Histochem Cell Biol, 2004, 121: 429~439.
[82]Chignola R. Self-potentiation of ligand-toxin conjugates containing ricin A chain fused with vira l structures. J Bio Chem, 1995,207: 23345~23351.
[83]Chairat T, Thawomchai L, Wiyada W, et al. Brit J Haenatol 2006,132(3): 370~373.
[84] Marsden CJ, Knight S, Smith DC, Day PJ, Roberts LM, Phillips GJ, et al. Insertional mutagenesis of ricinAchain: a novel route to an anti-ricin vaccine. Vaccine 2004;22(21–22):2800–5.and ricin to surface receptors of human cells. J Biol Chem, 1976, 251: 3977~3984.
[85]Riccobono F., Fiani M. L. Mannose receptor dependent uptake of ricin A1 and A2 chains by macrophage Carbohydr Res,1996, 282(2): 285~292.
[86]Dyba M, Tarasova NI, Michejda CJ. Small molecule toxins targeting tumor receptors. Curr Pharm Des 2004; 10: 2311–2334.
[87]Vallera DA et al. Targeting urokinase-type plasminogen activator receptor on human glioblastoma tumors with diphtheria toxin fusion protein DTAT. J Natl Cancer Inst 2002;94: 597–606.
[88]Morokoff AP, Novak U. Targeted therapy for malignant gliomas. J Clin Neurosci 2004; 11: 807–818.
[89]Frankel AE et al. Immunotoxin therapy of hematologic malignancies. Semin Oncol 2003; 30: 545–557.
[90]Chovnick A et al. A recombinant, membrane-acting immunotoxin. Cancer Res 1991; 51: 465–467.
[91]Choo AB, Dunn RD, Broady KW, Raison RL. Soluble expression of a functional recombinant cytolytic immunotoxin in insect cells. Protein Exp Purif 2002; 24: 338–347.
[92]Gasanov SE, Rael ED, Gasanov NE, Vernon LP. In vitro evaluation of Pyrularia thionin-anti-CD5 immunotoxin. Cancer Immunol Immunother 1995; 41: 122–128.
[93]Hinman CL, Tang HP. A membrane-lytic immunoconjugate selective for human tumor T-lymphocytes. Int J Immunopharmacol 1998; 20: 467–478.
[94]LeMaistre CF et al. Phase I trial of a ligand fusion-protein (DAB389IL-2) in lymphomas expressing the receptor for interleukin-2. Blood 1998; 91: 399–405.
[95]Frankel AE et al. A phase II study of DT fusion protein denileukin diftitox in patients with fludarabine-refractory chronic lymphocytic leukemia. Clin Cancer Res 2003; 9:3555–3561.
[96] Johan N.G et al. Phase II study of denileukin diftitox for relapsed/refractory B-Cell non-Hodgkin’s lymphoma. J Clin Oncol 2002; 22: 4095–4102.
[97]Khan T, Waring P. Macrophage adherence prevents apoptosis induced by ricin. Eur J Cell Bio l, 1993, 62: 406~414.
[98]江渝.程序型细胞死亡的生物化学基础.国外医学临床生物化学与检验学分册, 1996, 17: 178~182.
[99]Benis J. Signal transduction via the MAP kinase: proceed at your own RSK. Proc Natl Acad Sci USA, 1993, 91: 5889~5992.
[100]LinJY.Abrinandricin:Newanti-tumoursubstances.Nature ,1970;227(1):292.
[101]Sandra W Key, et al. Researchers know beans about cancer research. Cancer Weekly Plus, 1998,11: 20.
[102]Fred Wilson. Immunotoxin safely eliminates local inflammation in two days. Dermatology Times, 2000,21: 18~19.
[103]Weiner L M. Phase I evaluation of an antibreast carcinoma monoclonal antibody 260F9 ricin A chain. Cancer Res., 1989,49: 4062~4067.
[104]董巨莹,等.蓖麻毒素及其修饰物对U937细胞IL21B和TNF2A的诱导作用.细胞与分子免疫学杂志, 1999,15:123~125.
[105]张会.包有蓖麻毒素脂质体的制备、性质及毒性研究.生物化学与生物物理学报, 1999,31: 472~474.
[106]Chignola R. Self-potentiation of ligand-toxin conjugates containing ricin A chain fused with vira l structures. J Bio Chem, 1995,207: 23345~23351.
[107]Lord J. Ricin: structure, mode of action, and some current applications. FASEB J., 1994,8: 201~208.
[108]Frankel AE et al. Phase I trial of a novel diphtheria toxin/granulocyte macrophage colony-stimulating factor fusion protein (DT388GMCSF) for refractory or relapsed acute myeloid leukemia. Clin Cancer Res 2002; 8: 1004–1013.
[109]Weber F et al. Safety, tolerability, and tumor response of IL4-Pseudomonas exotoxin (NBI-3001) in patients with recurrent malignant glioma. J Neurooncol 2003; 64: 125–137.
[110]Rand RW et al. Intratumoral administration of recombinant circularly permuted interleukin-4-Pseudomonas exotoxin in patients with high-grade glioma. Clin Cancer Res 2000; 6:2157–2165.
[111]Sandvig K, van Deurs B. Entry of ricin and Shiga toxin into cells: molecular mechanisms and medical perspectives. EMBO J 2000; 19: 5943–5950.
[112]Johannes L, Goud B. Surfing on a retrograde wave: how does Shiga toxin reach the endoplasmic reticulum? Trends Cell Biol 1998; 8: 158–162.
[113]Simpson JC et al. Ricin A chain utilises the endoplasmic reticulum-associated protein degradation pathway to enter the cytosol of yeast. FEBS Lett 1999; 459: 80–84.
[114]Wesche J, Rapak A, Olsnes S. Dependence of ricin toxicity on translocation of the toxin A-chain from the endoplasmic reticulum to the cytosol. J Biol Chem 1999; 274: 34443–34449.
[115]Lord J. M et al. Retrograde transport of toxins across the endoplasmic reticulum membrane. Biochem Soc Trans 2003; 31:1260–1262.
[116]Mallard F, Johannes L. Shiga toxin B-subunit as a tool to study retrograde transport. In: Philpott D, Ebel F (eds). Methods in Molecular Medicine. Shiga Toxin Methods and Protocols,Vol 73, Chapter 17. Humana Press: Totowa, New Jersey, 2002,pp 209–220.
[117]Mallard F et al. Early/recycling endosomes-to-TGN transport involves two SNARE complexes and a Rab6 isoform. J Cell Biol 2002; 156: 653–664.
[118]Tai G et al. Participation of syntaxin 5/Ykt6/GS28/GS15 SNARE complex in transport from the early/recycling endosome to the TGN. Mol Biol Cell 2004; 15: 4011–4022.
[119]Natarajan R, Linstedt AD. A cycling cis-Golgi protein mediates endosome-to-Golgi traffic. Mol Biol Cell 2004; 15: 4798–4806.
[120]Lu L, Tai G, HongW. Autoantigen Golgin-97, an effector of Arl1 GTPase, participates in traffic from the endosome to the trans-Golgi network. Mol Biol Cell 2004; 15: 4426–4443.
[121]Wolf AA, Fujinaga Y, Lencer WI. Uncoupling of the cholera toxin-G(M1) ganglioside receptor complex from endocytosis, retrograde Golgi trafficking, and downstream signal transduction by depletion of membrane cholesterol. J BiolChem 2002;277: 16249–16256.
[121]Ramegowda B, Tesh VL. Differentiation-associated toxin receptor modulation,cytokine production, and sensitivity to Shiga-like toxins in human monocytes and monocytic cell lines. Infect Immun 1996; 64: 1173–1180.
[122]Tesh VL, Ramegowda B, Samuel JE. Purified Shiga-like toxins induce expression of proinflammatory cytokines from murine peritoneal macrophages. Infect Immun 1994; 62: 5085–5094.
[123]Hoey DE et al. Verotoxin 1 binding to intestinal crypt epithelial cells results in localization to lysosomes and abrogation of toxicity. Cell Microbiol 2003; 5: 85–97.
[124]Llorente A et al. Expression of mutant dynamin inhibits toxicity and transport of endocytosed ricin to the Golgi apparatus. J Cell Biol 1998; 140: 553–563.
[125]Grimmer S, Iversen TG, van Deurs B, Sandvig K. Endosome to Golgi transport of ricin is regulated by cholesterol. Mol Biol Cell 2000; 11: 4205–4216.
[126] Day P. J., Owens S. R., Wesche J., et al. An interaction between ricin and calreticulin that may have implications for toxin trafficking. J. Biol. Chem. 2001, 276: 7202~7208.
[127]Lauvrak SU, Llorente A, Iversen TG, Sandvig K. Selective regulation of the Rab9-independent transport of ricin to the Golgi apparatus by calcium. J Cell Sci 2002; 115: 3449–3456.
[128]Birkeli KA et al. Endosome-to-Golgi transport is regulated by protein kinase A type II alpha. J Biol Chem 2003; 278: 1991–1997.
[129]Teter K., Holmes R.K. Inhibition of endoplasmic reticulumassociated degradation in CHO cells resistant to cholera toxin, Pseudomonas aeruginosa exotoxin A, and ricin. Infect. Immun. 2002,70: 6172~6179.
[130]Pelkmans L, Burli T, Zerial M, Helenius A. Caveolin-stabilized membrane domains as multifunctional transport and sorting devices in endocytic membrane traffic. Cell 2004; 118: 767–780.
[131]Pelham HR. The dynamic organisation of the secretory pathway. Cell StructFunct 1996; 21: 413–419.
[132]Cosson P, Letourneur F. Coatomer (COPI)-coated vesicles: role in intracellular transport and protein sorting. Curr Opin Cell Biol 1997; 9: 484–487.
[133]Johannes L, Tenza D, Antony C, Goud B. Retrograde transport of KDEL-bearing B-fragment of Shiga toxin. J Biol Chem 1997; 272: 19554–19561.
[134]Doan, L.G., 2004. Ricin: mechanism of toxicity, clinical manifestations, and vaccine development.Areview. J. Toxicol.,Clin. Toxicol. 42, 201.
[135]Tracey K J , Beutler B, Lowry S F, et al. Shock and tissue injury induced by recombinant human cachetin. Science, 234: 470~474
[136]Griffiths, G.D., Newman, H..1986. Identification and quan-tification of ricin toxin in animal tissues using ELISA. J. Forensic Sci. Soc. 26, 349.
[137]Licastro F, Morini M C, Bolognesi A , et al. Ricin induced the production of tumor necrosis facto ralpha and interleuk in 21 beta by human peripheral2blood mononuclear cells. Biochem J , 1993, 294: 517~520.
[138]Muldoon D F, Bagchi D, Hassoun E A , et al. Themodulating effects of tumo r necro sis factor alpha antibody on ricin-induced oxidative stress in mice. J Biochem Toxico, 1994, 9: 311~318.
[139]董巨莹,王剑波,王文学.蓖麻毒素诱导小鼠肝脏产生肿瘤坏死因子.第四军医大学学报, 1997, 18: 78.
[140]Hassoun, E.A., Wang, X., Time- and concentration-dependent production of superoxide anion, nitric oxide, DNA damage and cellular death by ricin in the J774A. macrophage cells. J. Biochem. Mol. Toxicol. 1999, 13, 179.
[141]S.H. Pincus, T.K. Marcotte, B.M. Forsyth, H. Fang, In vivo testing of anti-HIV immunotoxins, Methods Mol. Biol. 2001, 166: 277–294.
[142]J. Zhan, Y. Chen, K. Wang, S. Zheng, Expression of ricin A chain and ricin A chain-KDEL in Escherichia coli, Protein Expr. Purif. 2004, 34:197–201.
[143]A. Miyawaki, Green fluorescent protein-like proteins in reef Anthozoa animals, Cell Struct. Funct. 2002, 27 :343–347.
[144]M. Chalfie, Y. Tu, G. Euskirchen, W.W. Ward, D.CGreen fluorescent protein as amarker for gene expression, Science 1994, 263: 802–805.
[145]MuldoonDF.Stohes S J. Ricin-induced oxidative stress in mice.Toxico logist,1991,11: 214.
[146]K. Sandvig, O. Garred, K. Prydz, J.V. Kozlov, S.H. Hansen, B. van Deurs, Retrograde transport of endocytosed Shiga toxin to the endoplasmic reticulum, Nature 1992,358:510–512.
[147]K. Sandvig, B. van Deurs, Transport of protein toxins into cells: pathways used by ricin, cholera toxin and Shiga toxin, FEBS Lett. 2002, 52: 49–53.
[148]Suzuki A, Doi H, Matsuzawa F, et al.Bcl-2 antiapoptotic protein mediates verotoxin II-induced cell death: possible association between bcl-2 and tissue failure by E. coli O157:H7. Genes Dev., 2000, 14: 1734～1740.
[149]Leek M D, Griffiths G D, Green M A. Intestinal patho logy follow ing intramuscular ricin poisoning. J Patho l, 1989, 159: 329~334.
[150]Waring P. DNA fragmentation induced in macrophages by gliotoxin does not require protein synthesis and is preceded raised ino sitotripho sphate levels. J Bio l Chem, 1990, 265: 14476~14480
[151]Olsnes S. The history of ricin, abrin and related toxins. Toxicon 2004;44:361–70.
[152]Vitetta ES, Thorpe PE. Immunotoxins containing ricin or it’s A chain. Semin Cell Biol 1991;2:47–58.
[153]Sadani GR, Soman CS, Deodhar K, Nadkarni GD. Reactive oxygen species involvement in ricin-induced thyroid toxicity in rat. Hum Exp Toxicol 1997;16:254–6.
[154]Moran, J. L., Siegel, D., Sun, X-M., Ross, D., 1996. Induction of apoptosis by benzene metabolites in HL-60 and CD34+ human bone marrow progenitor cells. Mol. Pharm. 50, 610-615
[155]Simon HU, Haj-Yehia A, Levi-Schaffer F. 2000. Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 5: 415-418.
[156]Gupta, A., Rosenberger, S. F., Bowden, G.T., 1999. Increased ROS levelscontribute to elevated transcription factor and MAP kinase activities in malignantly progressed mouse keratinocyte cell lines. Carcinogenesis 20, 2063-2073.
[157]Klotz, L. O., Pellieux, C., Briviba, K., Piierlot, C., Aubry, J. M., Sies, H., 1999. Mitogen-activated protein kinase (p38-, JNK-, ERK-)activation pattern induced by extracellular and intracellular singlet oxygen and UVA, Eur. J. Biochem. 260, 917-922.
[158]MacFarlane M, Cohen GM, Dickens M. 2000. JNK and p38 activation in receptor-mediated and chemically induced apoptosis of T-cells: differential requirements for caspase activation. Biochem J. 348:93-101.
[159]Werlen, G., Hausmann, B., Naeher, D., Palmer, E., 2003. Signaling life and death in the 0thymus: timing is everything. Science 299, 1859–1863.
[160]Halliwell B. Antioxidant defence mechanisms: from the beginning to the end (of the beginning). Free Radic Res 1999;31:261–72.
[161]Thannickal VJ, Fanburg BL. Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol 2000;279:L1005–28.
[162]Lewis TS, Shapiro PS, Ahn NG. Signal transduction through MAP kinase cascades. Adv Cancer Res 1998;74:49–139.
[163]Zhang W, Liu HT. MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res 2002;12:9–18.
[164]Ishikawa Y, Kitamura M. Dual potential of extracellular signal-regulated kinase for the control of cell survival. Biochem Biophys Res Commun 1999;264:696–701.
[165]Tibbles LA, Woodgett JR. The stress-activated protein kinase pathways. Cell Mol Life Sci 1999;55:1230–54.
[166]MacFarlane M, Cohen GM, Dickens M. JNK (c-Jun N-terminal kinase) and p38 activation in receptor-mediated and chemically induced apoptosis of T-cells: differential requirements for caspase activation. Biochem J 2000;348(Pt1):93–101.
[167]Obata T, Brown GE, Yaffe MB. MAP kinase pathways activated by stress: thep38 MAPK pathway. Crit Care Med 2000;28(Suppl. 4):N67–77.
[168]Gupta S, Barrett T, Whitmarsh AJ, Cavanagh J, Sluss HK, Derijard B, et al. Selective interaction of JNK protein kinase isoforms with transcription factors. EMBO J1996;15:2760–70.
[169]Jiang Y, Gram H, Zhao M, New L, Gu J, Feng L, et al. Characterization of the structure and function of the fourth member of p38 group mitogen-activated protein kinases,p38delta. J Biol Chem 1997;272:30122–8.
[170]Kang CD, Yoo SD, Hwang BW, Kim KW, Kim DW, Kim CM, et al. The inhibition of ERK/MAPK not the activation of JNK/SAPK is primarily required to induce apoptosis in chronic myelogenous leukemic K562 cells. Leuk Res 2000;24:527–34.
[171]Anderson SM, Reyland ME, Hunter S, Deisher LM, Barzen KA,Quissell DO. Etoposide-induced activation of c-jun N-terminal kinase (JNK) correlates with drug-induced apoptosis in salivary gland acinar cells. Cell Death Differ 1990;6:454–62.
[172]Quissell DO, Barzen KA, Gruenert DC, Redman RS, Camden JM, Turner JT. Development and characterization of SV40 immortalized rat submandibular acinar cell lines. In Vitro Cell Dev Biol Anim 1997;33:164–73.
[173]Wang X, Martindale JL, Liu Y, Holbrook NJ. The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 1998;333(Pt 2):291–300.
[174]Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–5.
[175]Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979;76:4350–4.
[176]Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel AR. PD098059 is a specific inhibitor of the activation of mitogen-activated protein kinase in vitro and in vivo. J Biol Chem 1995;270:27489–94.
[177]Bennett BL, Sasaki DT, Murray BW, O’Leary EC, Sakata ST, Xu W, et al. SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci USA 2001;98:13681–6.
[178]Davies SP, Reddy H, Caivano M, Cohen P. Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 2000;351:95–105.
[179]Kyaw M, Yoshizumi M, Tsuchiya K, Kirima K, Suzaki Y, Abe S, et al. Antioxidants inhibit endothelin-1 (1–31)-induced proliferation of vascular smooth muscle cells via the inhibition of mitogen-activated protein (MAP) kinase and activator protein-1 (AP-1). Biochem Pharmacol 2002;64:1521–31.
[180]Martindale JL, Holbrook NJ. Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol 2002;192:1–15.
[181]Wang H, Joseph JA. Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 1999;27:612–6.
[182]Filomeni G, Rotilio G, Ciriolo MR. Disulfide relays and phosphorylative cascades: partners in redox-mediated signaling pathways. Cell Death Differ 2005;12:1555–63.
[183]Caughman GB, Schuster GS, Rueggeberg FA. Cell lipid alterations resulting from prolonged exposure to dimethylaminoethylmethacrylate. Clin Oral Investig 1999;3:181–7.
[184]Bereznowski Z. Effect of methyl methacrylate on mitochondrial function and structure. Int J Biochem 1994;26:1119–27.
[185]BoguskiM S, McintoshM W. Biomedical informations for p roteomics〔J〕. Nature, 2003, 422 (6928) : 233 - 237.
[186]Petricoin E , Ardekani A M, Hitt B A , et al. Use of proteomic patterns in serum to identify ovarian cancer[J ] . The Lancet , 2002 , 359 :572
[187]Rabilloud T. Two-dimensional gel electrophoresis in proteo-mics: old, old fashioned, but is still climbs up the moun-tains[J].Proteomics, 2002,2(1):3-10.
[188]Davidsson P, Paulson L, Hesse C, Blennow K, NilssonCL. Proteome studies ofhuman cerebrospinal fluid and brain tissue using a preparative two-dimensional electrophoresis approach prior to mass spectrometry [J]. Proteomics, 2001 (3):444-452.
[189]WilkinsM R, Sanchez J C, Gooley A A, et al. Progresswith p roteome p rojects: why all proteins exp ressed by a genome should be identified and how to do i〔tJ〕. Biotechnology and genetic engineering reviews, 1996, 13: 19 - 50.
[190]李义良,朱华清等.双向电泳技术〔J〕.生物学教学, 2004, 29 (4) : 64 - 66.
[191]Stocks J . Miller NE. Analysis of high density lipopmtein apoli poproeins by eapillary zone and capillary SDS gel electropharesis J Lipid Res 1998 Jan ;39 (1) :218 - 227.
[192]Leeds J. A, Beck with J.Lambd are pressor N 2 terminal DNA binding domain as an assay for protein transmembrane segment interaction in vivo[J].JMolBiol,1998, 280(5) :7992810
[193]Rigaut G, ShevchenkoA, RutzB, et al.A generic protein purification method for protein complex characterization and proteome exploration[J].Nat Biotechnol, 1999, 17 (10 ) :21-32
[194]Krogan N, Cagney G,Yu H, et al.Global land scape of protein Complexes in the yeast Saccharomycesce revisiae [J]. Nature, 2006, 440(7084) : 6372643
[195]Schubert W, Bonnekoh B, Pommer A. J, et al. Analyzing proteome to pology and function by automated multidimension alfluorescence microscopy [J]. Nat Biotechnol, 2006, 24(10) :127021278
[196]TastoJ J, Carnahan R H, Mc Donald W. H, et al. Vectors and gene targeting modules for tandem affinity purificationin Schizosaccharomycespombe [J].Yeast,2001, 18(7) :6572662
[197]Tasto J. J, Carnahan R. H, McDonald W. H, et al. Vectors and gene targeting modules for tandem affinity purificationin Schizosac charomy cespombe [J].Yeast,2001, 18 (7) :6572662
[198]Hu C. D, Chinenov Y, Kerppola T. K.Visualization of interactions among bZip and Rel family protein sinliving cell susing bimolecular Fluorescencecomplementation [J].MolCell, 2002, 9(4) :7892798
[199]Morell M, Espargaro A, Aviles F, et al. Detection of transient protein–protein interactions by bimolecular fluorescence complementation: the Abl 2SH3 case [J].Proteomics,2007,7 (7) :21-36
[200]Grinberg A. V, Hu C. D, Kerppola T. K.Visualization of Myc PmaxP Mad family dimers and the competition for dimerization in living cells [J].Mol Cell Biol, 2004, 24 (10) : 424-308
[201]Fuentes M, Mateo C, Pessela B. C, et al. Solid phase proteomics: Dramatic reinforcement of very weak protein–protein interactions [J].J Chromatogr B Analyt Technol Biomed Life Sci, 2007, 849 (122) :243-250
[202]Zeghouf M, Li J, Butland G, et al. Sequential peptide affinity (SPA) system for the identification of mammalian and bacterial proteincomplexes [J].J ProteomeRes,2004, 3 (3) :4632468
[203]Cheeseman I. M, Desai A. A. combined approach for the localization and tandem affinity purification of protein complexes from metazoans [J].Sci STKE, 2005, 2005(266) :pl1
[204]Burckstummer T, Bennett K. L, PreradovicA, et al. An efficient tandem affinity purification procedure for interaction proteomics in mammalian cells [J].Nat Methods, 2006, 3 (12) :1013-1019
[205]Rubio V, Shen Y, SaijoY, et al. An alter native tandem affinity purification strategy applied to Arabidopsis protein complexisolation [J]. Plant J, 2005, 41(5) :767-778
[206]孙宇,贾凌云,任军.蛋白质相互作用的研究方法[J].分析化学2007, 35(5) :760-766
[207]Washburn M P, Wolters D, YatesIII J R. Large-scale analysis of the yeast proteome by multi-dimensional protein identification technology [J]. Nat Biotechnol, 2001, 19 (3): 242-247.
[208]Florens L, Vashburn M P.Proteomic analysis by multidimensional protein identification technology [J].Methods Mo1 Bio1, 2006,328:159-175.
[209]Maor R, Jones A, Thomas S, et al.Multidimensional protein identification technology analysis of ubiquitinated proteins in plants [J]. Mol Cell Proteomics, 2007, 6: 601-610.
[210]Rabilloud T.Two-dimensional gel electrophoresis in proteomics: old, old fashioned, but it still climbs up the mountains [J].Proteomics, 2002, 2: 3-10.
[211]Corpet F, Servant F, Gouzy J, et al.ProDom and ProDom-CG:Tools for protein domain analysis and whole genome comparisons [J] . Nucleic Acid Res, 2000,28:267-269.
[212]Salagado H, Moreno-Hagelsieb G, Smith T F, et al. Operons in Escherichia coli: Genomic analyses and predictions[J] . Proc Natl Acad Sci USA, 2000,97(12):6652-6657.
[213]Snel B, Lehmann G, Bork P, et al.A web-server to retrieve and display the repeatedly occurring neighbourhood of a gene [J] . Nucleic Acids Res, 2000,28:3442-3444.
[214]Cruzado ID , Coekrill SL. MeNeal CJ , et al . Charactenzation and quanti-tation of apolipoprotein B - 100 by capillary electrophoresis. J Lipid Bes 1998 Jan ; 39(1) ;205 - 217.
[215]TERABE S , OTSU KA K.et al. Elect rokinetic separation with micellar solutions and open tuber capillaries [J ] . Anal Chem , 1984 , 56 : 111 2113.
[216]Hjierten BH , Ceriotti L. Shibata T , et al . Analysis of lipoproteins by eapillary zone electrophoresis in microfluidie devices : assay develop -ment and surtace roughness measurements. Anal Chem. 2002 Apr 1 ;74(7) :1702 - 11.
[217]Cohen G. A , Kanger, Athias A , Persegol L , Lagrost L , gambert P , Lallemant C.Capillary gel electrophoresis analysis of apolipoproteins A - I and A– 11 in human high - density lipoproteins. Anal Biochem. 1994 May 1 ; 218 (2) :320 - 4.
[218]Proctor SD , Mama JC Separation and quantifieation of apolipopro– tein B - 48 and other apolipoproteins by dynamic sieving capillary e - lec2 trophoresis. J Lipid Res. 1997 Feb ; 38 (2) :410 - 414.
[219]Rodriguez Diaz R , Zhu M.et al J Chromatogr A 1997 ;772 :145 - 160.
[220]Stocks J ,Miller NE. Analysis of apolpoproteins and lipopmteins by eapillary electrophoresis Electrophoresis 1999 Jul ; 20 (10) :2118 - 23.
[221]Sehmitz G, Vollers C , Richter V. Analytical capillary isota - ehophoresis; a routine technique for the analysis of lipoproteins and lipoprotein subfractions in whole serum. J Chromalogr. 1985 ;320 :253- 262.
[222]Jones D T, Swindells M B.Getting the most from PSI-BLAST [J].Trends Biochem Sci, 2002, 27:161-164.
[223]史志诚动物毒物学中国农业出版社[M], 2001,394-396
[224]赵英蓖麻毒素毒性作用机理研究进展中华预防医学杂志1999;33(1):54-56
[225]Rita M. et al. Identification of three oligosaccharide binding sites in Ricin[J]. Biochemistry, 1999,38: 11677-11685.
[226]曾佑炜蓖麻毒蛋白的分离纯化和毒理作用研究2004;20(4):23-25
[227]SPARANO A, LATHERS D M, et al. Modulation of Th1 and Th2 cytokine profiles and their association with advanced head and neck squamous cell carcinoma [ J].Otolaryngol Head Neck Surg, 2004, 131( 5) :573- 576.
[228]AGARWAL A, RANI M, SAHA G K, et al. Disregulated expression of the Th2 cytokine gene in patients with intraoral squamous cell carcinoma [ J] . Immunol Invest, 2003, 32: 17- 30.
[229]HESS A D, THOBURN C,.et al. The N-terminal flanking region of the invariant chain peptide augments the immunogenicity of a cryptic“self”epitope from a tumorassociated antigen [ J] . Clin Immunol, 2001, 101( 1) : 67- 76.
[230]Battelli, M.G., Musiani, S., Buonamici, L., Santi, S., Riccio, M., Maraldi,N.M., Girbes, T., Stirpe, F.. Interaction of volkensin with HeLa cells: binding, uptake, intracellular localization, degradation and exocytosis. Cell. Mol. Life Sci. ,2004,61:1975– 1984.
[231]Bolognesi, A., Polito, L..Immunotoxins and other conjugates: preclinical studies. Mini Rev. Med. Chem. 2004,4:563– 583.
[232]Bradberry, S.M., Dickers, K.J., Rice, P., Griffiths, G.D., Vale, J.A.. Ricinpoisoning. Toxicol. Rev. 2003,22:65– 70.
[233]Brown, R.F., White, D.E.. Ultrastructure of rat lung following inhalation of ricin aerosol. Int. J. Exp. Pathol. ,1997,78:67– 276.
[234]Pryme, I.F., Bardocz, S., Pusztai, A., Ewen, S.W.. Dietary mistletoe lectin supplementation and reduced growth of a murine non-Hodgkin lymphoma.Histol.Histopathol. 2002,17:261– 271.
[235]Citores, L., Mun?oz, R., Rojo, M.A., Jime′nez, P., Ferreras, J.M., Girbe′s, T. Evidence for distinct cellular internalisation pathways of ricin and nigrin b. Cell. Mol. Biol.2003,49:461465.
[236]Kreitman,R.J.Immunotoxins.ExpertOpin.Pharmacother. 2000,1:1117–1129.
[237]Doan, L.G. Ricin: mechanism of toxicity, clinical manifestations,and vaccine development. A review. J. Toxicol. Clin. Toxicol.2004, 42:201-208.
[238]Fracasso, G., Bellisola, G., Castelletti, D., Tridente, G., Colombatti, M. Immunotoxins and other conjugates: preparation and general characteristics. Mini Rev.Med. Chem.2004,4:545–562.
[239]Frank, S.A., Nowak, M.A. Developmental predisposition to cancer. Nature. 2003,4:42, 494.
[240]Kyriakis JM,Avruch J. Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation[J]. Physiol Rev,2001, 81(2):807-869.
[241]MU Jin-ye,CHENXIao-Guang.Progress of the study on the mitogen activated protein kinase pathways [J].Chin Bull Life Sci,2002,14(4):208-211.
[242]Ghatan S,Larner S,Kinoshita Y,et al. p38 MAP kinase mediates bax translocation in nitric oxide-induced apoptosis in neurons[J].JCell Biol, 2000,150(2):335~347.
[243]Barancik M,Bohacova V,Kvackajova J,et al. SB203580, a specific inhibitor of p38-MAPK pathway, is a new reversal agent of P-glycoprotein-mediated multidrug resistance [J].Eur J Pharm Sci,2001,14(1):29~36.
[244] Brewster J. L, De Valoir T,Dwyer N. C et al.An osmosensing signaltransduction pathway in yeast [J].Science, 1993, 259(5102):1760~1763.
[245] Han J, Lee J. D, Bibbs L,et al. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells [J]. Science, 1994, 265(5173):808~811.
[246] Obata T, Brown GE, Yaffe MB. MAP kinase pathways activated by stress:the p38 MAPK pathway [J].Crit Care Med,2000,28(1):67~77.
[247] Ketam S,John F,Brion N,et al.Inhibition of p38 mitogen-activated protein kinase increases LPS induced inhibition of apoptosis in neutrophils by activating extracellular signal regulated kinase [J].Surgery,2001,130(2):242~247.
[248] Wang XS,Diener K,Manthey CL,et al, Molecular cloning and characterization of a novel p38 mitogen-activated protein kinase [J].J Biol Chem,1997,272(38):23 668~23 674.
[249] Boder AJ.Ye R,Lees-Miller SP.Uv-light induces p38MAPK-dependent phosphorylation of Bel10 [J].Biochem Biophys Res Commun,2003, 301(4): 923~926.
[250] Heyse S, Vogel H, Sanger M, Sigrist H. Covalent attachment of functionalised lipid bilayers to planar waveguides for measuring protein binding to biomimetic membranes. Protein Sci 1995;4:2532–44.
[251]黄丽俊等.蛋白质组研究技术及其进展〔J〕.生物学通报, 2005, 40 (8) : 4-7.
[252]WilkinsM R, Sanchez J C, Gooley A A, et al. Progresswith p roteome p rojects: why all p roteins exp ressed by a genome should be identified and how to do i〔tJ〕. Biotechnology and genetic engineering reviews, 1996, 13: 19~50.
[253]吴世容,李志良,李根容,等.生物质谱的研究及其应用.重庆大学学报, 2004, 27(1):123~127.
[254]成海平,钱小红.蛋白质组研究的技术体系及其进展.生物化学与生物物理进展,2000,27:584~588.
[255]Lay. JOJr.MALDI-TOF mass spectrometry of bacteria. [J]. MassSpectrom Rev, 2001; 20(4):172~194.
[256]HarveyDJ.Identification of protein-bound carbohydrates by masss pectrometry [J]. Proteomic, 2001; 1(2):311~328.
[257]刘慧敏,赖志辉,黎军,等.碎片结构分析在MALDIT OF MS法测定多肽序列中的应用.生物化学与生物物理学报, 2000, 32:179~182.
[258]Ong S E, MannM. Mass spectrometry based p roteomics turns quantitative〔J〕. Nat ChemBiol, 2005, 1 (5) : 252 ~257.
[259]Somiari RI, Somiari S, Russell S, et al. Proteomics of breast carcinoma [J]. Chromatogr B Analyt Technol Biomed Life Sci, 2005,815(1- 2):215~225.
[260]Malik G,Ward MD,Gupta SK,et al. Serum levels of an isoform of apolipoprotein A-Ⅱas a potential marker for prostate cancer[J]. Clin Cancer Res, 2005, 11(3):1073~1085.
[261]Eggeling FV,Davies H,Lomas L, et al. Tissue-specific microdissection coupled with ProteinChip array technologies applications in cancer research[J]. Biotechniques, 2000, 29(5): 1066~1070.
[262]Landuyt B, Prenen H,Debiec-Rychter M,et a1. Differential protein expression profile in gastrointestinal stromal tumors[J]. Amino Acids, 2004, 27: 335~337.
