全反式视黄酸诱导小鼠腭裂畸形血清比较蛋白质组学研究
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摘要
目的使用全反式视黄酸(all-trans retinoic acid,atRA)诱导腭发育敏感期的小鼠作为模型,利用血清蛋白质组学技术,对血清中差异表达的蛋白质和多肽进行鉴定和验证,探讨腭裂发病机制。
方法60只C57BL/6J系孕鼠,妊娠10天给予全反式视黄酸(100 mg/kg),建立小鼠腭裂模型,采集妊娠14天孕鼠血清。实验分为三组：全反式视黄酸组、空白组、植物油对照组。采用双向电泳(2-DE)、偶联基质辅助激光解吸电离—飞行时间质谱(MALDI-TOF MS)技术,寻找腭裂孕鼠血清差异表达蛋白质。并通过Western blot对试验结果进行验证。
结果触珠蛋白β-chain(Haptoglobinβ-chain, Hpβ-chain)和血清淀粉样P成分(Serum amyloi d P-component, SAP)在全反式视黄酸诱导腭裂孕鼠血清中表达上调,分别是正常孕鼠血清蛋白含量的2.5和3倍。另外还发现触珠蛋白β-chain在全反式视黄酸组上调表达的同时发生糖基化修饰的变化。Westernblot验证结果与2-DE结果相一致。
结论HPβ-chain和SAP在全反式视黄酸致腭裂孕鼠血清中表达上调,为进一步研究全反式视黄酸致小鼠腭裂畸形的病理机制打下基础。同时,发现HPβ-chain在腭裂孕鼠血清中糖基化修饰改变,提示糖基化修饰在全反式视黄酸致腭裂畸形中具有重要调控作用。
Objective To explore the protein changes in serum of the cleft palate disease, the animal model of the retinoic acid induced cleft palate and the comparative proteome analysis were used in this study.
Methods Sixty C57BL/6J strains of mice were used in this study. For the experiment group, E10 pregnant female mice were given by gastric intubations of a single dose of all-trans retinoic acid (RA) (100 mg/kg) and the RA was dissolved in olive oil. The control group were given the equivalent volume of the carrier alone. The normal group did not give anything. Serum were collected from the pregnant mice at E14.2-DE, MALDI-TOF MS, and Western blotting were applied to analysis the protein changes in the serum of the normal, control, and RA pregnant mice groups.
Results Haptoglobin B-chain and serum amyloid P-component levels in sera of RA pregnant mice were significantly higher than those from normal or control subjects.Volum% for Haptoglobin B-chain and serum amyloid P-component were 3 and 2.5 times higher,respectively. In addition, N-glycans of haptoglobin B-chain were found differently altered expression between RA groups and normal or control groups. Each of these differences was statistically significant(p value<0.01),p value obtained by Mann-Whitney U-test.
Conclusion The expression level of the protein, haptoglobinβ-chain and serum amyloid P-component, in the serum of RA pregnant mice group were significantly higher than those of the normal and control groups. Differences in structure of the N-glycans of haptoglobin B-chain were found in RA pregnant mice. It was suggested that these potential functional proteins provide insights into the pathogenetic mechanism of cleft palate caused by excess RA remains.
方法60只C57BL/6J系孕鼠,妊娠10天给予全反式视黄酸(100 mg/kg),建立小鼠腭裂模型,采集妊娠14天孕鼠血清。实验分为三组：全反式视黄酸组、空白组、植物油对照组。采用双向电泳(2-DE)、偶联基质辅助激光解吸电离—飞行时间质谱(MALDI-TOF MS)技术,寻找腭裂孕鼠血清差异表达蛋白质。并通过Western blot对试验结果进行验证。
结果触珠蛋白β-chain(Haptoglobinβ-chain, Hpβ-chain)和血清淀粉样P成分(Serum amyloi d P-component, SAP)在全反式视黄酸诱导腭裂孕鼠血清中表达上调,分别是正常孕鼠血清蛋白含量的2.5和3倍。另外还发现触珠蛋白β-chain在全反式视黄酸组上调表达的同时发生糖基化修饰的变化。Westernblot验证结果与2-DE结果相一致。
结论HPβ-chain和SAP在全反式视黄酸致腭裂孕鼠血清中表达上调,为进一步研究全反式视黄酸致小鼠腭裂畸形的病理机制打下基础。同时,发现HPβ-chain在腭裂孕鼠血清中糖基化修饰改变,提示糖基化修饰在全反式视黄酸致腭裂畸形中具有重要调控作用。
Objective To explore the protein changes in serum of the cleft palate disease, the animal model of the retinoic acid induced cleft palate and the comparative proteome analysis were used in this study.
Methods Sixty C57BL/6J strains of mice were used in this study. For the experiment group, E10 pregnant female mice were given by gastric intubations of a single dose of all-trans retinoic acid (RA) (100 mg/kg) and the RA was dissolved in olive oil. The control group were given the equivalent volume of the carrier alone. The normal group did not give anything. Serum were collected from the pregnant mice at E14.2-DE, MALDI-TOF MS, and Western blotting were applied to analysis the protein changes in the serum of the normal, control, and RA pregnant mice groups.
Results Haptoglobin B-chain and serum amyloid P-component levels in sera of RA pregnant mice were significantly higher than those from normal or control subjects.Volum% for Haptoglobin B-chain and serum amyloid P-component were 3 and 2.5 times higher,respectively. In addition, N-glycans of haptoglobin B-chain were found differently altered expression between RA groups and normal or control groups. Each of these differences was statistically significant(p value<0.01),p value obtained by Mann-Whitney U-test.
Conclusion The expression level of the protein, haptoglobinβ-chain and serum amyloid P-component, in the serum of RA pregnant mice group were significantly higher than those of the normal and control groups. Differences in structure of the N-glycans of haptoglobin B-chain were found in RA pregnant mice. It was suggested that these potential functional proteins provide insights into the pathogenetic mechanism of cleft palate caused by excess RA remains.
引文
[1]Mossey P, Castillia E. Global registry and database on craniofacial anomalies. Geneva:World Health Organization,2003:37-43.
[2]Jiang R, Bush JO, Lidral AC. Development of the upper lip:morphogenetic and molecular mechanisms. Dev Dyn,2006,235:1152-1166.
[3]Fraser FC. The genetics of cleft lip and palate:yet another look. In Current Research Trends in Prenatal Craniofacial Development, RM Pratt, RL Christiansen, eds. (New York, NY:Elsevier North Holland Inc),1980:357-366.
[4]Francesco Carinci, Luca Scapoli, Annalisa Palmieri, et al. Human genetic factors in nonsyndromic cleft lip and palate:An update. International Journal of Pediatric Otorhinolaryngology.2007,71:1509-1519.
[5]Song Jukun, Zhou Jinglin, SHi Bing, et al. A preliminary investigation on plasma of non-syndromic cleft lip and/or palate using nuclear magnetic resonance-based metabonomics. West China Journal of Stomatology.2009,27(2):147-153.
[6]Christine G, Wenbo Zhi, Bao-Ling Adam. Proteomic Identification of Serum Biomarkers for Head and Neck Cancer Surveillance. Laryngoscope.2009,119(7):1291-1302.
[7]Hofmann C and Eichele G. Biology, chemistry, and medicine.In:The Retinoids, edited by M. B. Sporn, A. B. Roberts, and D. S.Goodman. New York:Raven.1994,387-441.
[8]Ross, S.A., De Luca, L.M. et al. Retinoids in embryonal development. Physiol. Rev.2000,80: 1021-1054.
[9]Kochhar, D.M., Johnson, E.M. Morphological and autoradiographic studies of cleft palate induced in rat embryos by maternal hypervitaminosis A. J. Embryol. Exp. Morphol.1965,14: 223-238.
[10]Abbott, B.D., Harris, M.W., Birnbaum, L.S. Etiology of retinoic acidinduced cleft palate varies with the embryonic stage. Teratology.1989,40:533-553.
[11]Kochhar, D.M., Johnson. E.M. Morphological and autoradiographic studies of cleft palate induced in rat embryos by maternal hypervitaminosis A. J. Embryol. Exp. Morphol.1965,14: 223-238.
[12]Giroud, A.& Martinet, M. Teratogenese par hautes doses de vitamine A en fonction des stades du developpement. Archs. Anat. Microsc.1956,45:77-98.
[13]Junko Okano, Shigehiko Suzuki, Kohei Shiota. Involvement of apoptotic cell death and cell cycle perturbation in retinoicacid-induced cleft palate in mice. Toxicology and Applied Pharmacology.2007,221:42-56.
[14]Padmanabhan, R. and A. Ijaz. Retinoic acid-induced asymmetric craniofacial growth and cleft palate in the to mouse fetus. Reproductive Toxicology.1997,11(6):843-860.
[15]Okano, J., S. Suzuki, et al. Involvement of apoptotic cell death and cell cycle perturbation in retinoic acid-induced cleft palate in mice. Toxicology and Applied Pharmacology.2007,221(1): 42-56.
[16]Tseng, C. F., C. C. Lin, et al. Antioxidant role of human haptoglobin. PROTEOMICS.2004, 4 (8):2221-2228.
[17]Saso, L., P. Tommasino, et al. Changes of acute-phase proteins in streptozotocin-induced diabetic rats. Physiol Res.2000,49(4):403-9.
[18]Van Vlierberghe, H., M. Langlois, et al. Haptoglobin polymorphisms and iron homeostasis in health and in disease. Clinica Chimica Acta.2004,345(1-2):35-42.
[19]Bicho, M. C., A. Pereira da Silva, et al. Sex steroid hormones influence the risk for cervical cancer:modulation by haptoglobin genetic polymorphism. Cancer Genetics and Cytogenetics. 2000,191(2):85-89.
[20]Hamrita, B., K. Chahed, et al. Proteomics-based identification of [alpha]1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas. Clinica Chimica Acta.2009,404(2):111-118.
[21]Vanhooren, V., X.-E. Liu, et al. N-glycan profiles as tools in diagnosis of hepatocellular carcinoma and prediction of healthy human ageing. Mechanisms of Ageing and Development. 2008,130(1-2):92-97.
[22]Istvan Liko, Marianna Mak, et al. Evidence for an extended interacting surface between [beta]-amyloid and serum amyloid P component. Neuroscience Letters.2007,412(1):51-55.
[23]Song, Z., L. Cai, et al. Accumulation and expression of serum amyloid P component in human atherosclerotic lesions. Atherosclerosis In Press, Corrected Proof.
[24]Szilvia Veszelka, Zoltan Urbanyi, et al. Human serum amyloid P component attenuates the bacterial lipopolysaccharide-induced increase in blood-brain barrier permeability in mice. Neuroscience Letters.2003,352(1):57-60.
[25]Urbanyi, Z., Lajos Laszlo, et al. Serum amyloid P component induces neuronal apoptosis and [beta]-amyloid immunoreactivity. Brain Research.2003,988(1-2):69-77.
[26]Zoltan Urbanyi, Miklos Sass, et al.Serum amyloid P component induces TUNEL-positive nuclei in rat brain after intrahippocampal administration. Brain Research.2007,1145:221-226.
[27]Vamsidhara C Dhulipala. Cell cycle proteins in normal and chemicallyinduced abnormal secondary palatedevelopment:a review [J]. Human & Experimental Toxicology (2006) 25:675-682.
[28]李鑫,金岩等.细胞程序性死亡及抗凋亡基因bcl-2在C57BL6N系小鼠腭裂形成中的意义[J]现代预防医学.2007,34(5)：858-860.
[29]李杨,石冰.非综合征唇腭裂的候选基因位点.口腔颌面外科杂志.2007,17(1)：82-85
[30]王文恭,童坦君.细胞凋亡过程中bacp-2基因的甲基化。[J]中国生物化学与分子生物学报。1998,14(3)：309-313.
[31]王海燕,王启会.细胞凋亡的相关基因调控.现代预防医学.2007,34(5)858-860.王如,刘彬.腭发育不同时期全反式视黄酸对腭突细胞增殖和凋亡的影响.华西口腔医学杂志.2008,26(5)：546-549.
[32]李鑫,金岩.细胞凋亡及p53基因的表达在C57BL6N系小鼠腭裂形成中的意义.实用口腔医学杂志.2000,16(6)：440-442.
[33]Jiu-Zhen Jin and Jixiang Ding. Analysis of cell migration, transdifferentiation and apoptosisduring mouse secondary palate fusion. Development 2006,133:3341-3347
[34]Gregg Duester. Retinoic Acid Regulation of the Somitogenesis Clock. Birth Defects Research 2007,81:84-92
[35]Vamsidhara C Dhulipalal, Wade V Welshons. Cell cycle proteins in normal and chemically induced abnormal secondary palatedevelopment:a revieW Human & Experimental Toxicology 2006,25:675-68
[1]Aivaliotis, M., et al., Large-scale identification of N-terminal peptides in the halophilic archaea Halobacterium salinarum and Natronomonas pharaonis. Journal of Proteome Research,2007.6(6): p.2195-2204.
[2]Kuyama, H., et al., A method for N-terminal de novo sequence analysis of proteins by matrix-assisted laser desorption/ionization mass spectrometry. Analytical Biochemistry,2008. 380(2):p.291-296.
[3]Roemer, I., et al., Analysis of mouse beta-haptoglobin chain by lectin affinoblotting detection. Electrophoresis,2001.22(14):p.3038-3042.
[4]Golaz, O., et al., Rapid Detection of the Main Human Plasma Glycoproteins by 2-Dimensional Polyacrylamide-Gel Electrophoresis Lectin Affinoblotting. Electrophoresis,1995.16(7):p. 1187-1189.
[5]Marynen, P., et al., A Genetic-Polymorphism in a Functional Domain of Human-Pregnancy Zone Protein-the Bait Region-Genomic Structure of the Bait Domains of Human-Pregnancy Zone Protein and Alpha-2 Macroglobulin. Febs Letters,1990.262(2):p.349-352.
[6]Claesson, R., et al., A new cleavage site for elastase within the complement component 3. Apmis,2010.118(10):p.765-768.
[7]Horie, M., et al., The N-terminal region is important for the nuclease activity and thermostahility of the flap endonuclease-1 from Sulfolobus tokodaii. Bioscience Biotechnology and Biochemistry,2007.71(4):p.855-865.
[8]Huang, Z.H., et al., Protein sequencing by matrix-assisted laser desorption ionization-postsource decay mass spectrometry analysis of the N-tris(2,4,6-trimethoxyphenyl)phosphine-acetylated tryptic digests. Analytical Biochemistry, 1999.268(2):p.305-317.
[9]An, M.R., et al., Efficient and clean charge derivatization of peptides for analysis by mass spectrometry. Rapid Communications in Mass Spectrometry,2010.24(13):p.1869-1874.
[10]Chamot-Rooke, J., et al., The combination of electron capture dissociation and fixed charge derivatization increases sequence coverage for O-glycosylated and O-phosphorylated peptides. Journal of the American Society for Mass Spectrometry,2007.18(8):p.1405-1413.
[11]Gallien, S., et al., Ortho-proteogenomics:Multiple proteomes investigation through orthology and a new MS-based protocol. Genome Research,2009.19(1):p.128-135.
[12]Franck, J., et al., On-Tissue N-Terminal Peptide Derivatizations for Enhancing Protein Identification in MALDI Mass Spectrometric Imaging Strategies. Analytical Chemistry,2009. 81(20):p.8305-8317.
[l]钱小红,贺福初.蛋白质组学：理论与方法.北京：科学出版社,2003.
[2]夏其昌,曾嵘.蛋白质化学与蛋白质组学.北京：科学出版社,2004.
[3]Wasinger, V. C., Cordwell, S. J., Cerpa-Poljak, A., Yan, J. X., Gooley, A. A., Wilkins, M. R., Duncan, M. W., Harris, R., Williams, K. L., and Humphery-Smith, I. (1995). Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis 16,1090-4.
[4]Klose, J. (1975). Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals. Humangenetik 26,231-43.
[5]O'Farrell, P. H. (1975). High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250,4007-21.
[6]Rabilloud, T., Chevallet, M., Luche, S., and Lelong, C. Two-dimensional gel electrophoresis in proteomics:Past, present and future. Journal of Proteomics 73,2064-2077.
[7]Karas, M., and Hillenkamp, F. (1988). Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. Anal Chem 60,2299-301.
[8]Gygi, S. P., Rist, B., Gerber, S. A., Turecek, F., Gelb, M. H., and Aebersold, R. (1999). Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat Biotechnol 17,994-9.
[9]Gygi, S. P., Rist, B., Griffin, T. J., Eng, J., and Aebersold, R. (2002). Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags. J Proteome Res 1,47-54.
[10]Tseng, C. F., Lin, C. C., Huang, H. Y., Liu, H. C., and Mao, S. J. T. (2004). Antioxidant role of human haptoglobin. PROTEOMICS 4,2221-2228.
[2]Jiang R, Bush JO, Lidral AC. Development of the upper lip:morphogenetic and molecular mechanisms. Dev Dyn,2006,235:1152-1166.
[3]Fraser FC. The genetics of cleft lip and palate:yet another look. In Current Research Trends in Prenatal Craniofacial Development, RM Pratt, RL Christiansen, eds. (New York, NY:Elsevier North Holland Inc),1980:357-366.
[4]Francesco Carinci, Luca Scapoli, Annalisa Palmieri, et al. Human genetic factors in nonsyndromic cleft lip and palate:An update. International Journal of Pediatric Otorhinolaryngology.2007,71:1509-1519.
[5]Song Jukun, Zhou Jinglin, SHi Bing, et al. A preliminary investigation on plasma of non-syndromic cleft lip and/or palate using nuclear magnetic resonance-based metabonomics. West China Journal of Stomatology.2009,27(2):147-153.
[6]Christine G, Wenbo Zhi, Bao-Ling Adam. Proteomic Identification of Serum Biomarkers for Head and Neck Cancer Surveillance. Laryngoscope.2009,119(7):1291-1302.
[7]Hofmann C and Eichele G. Biology, chemistry, and medicine.In:The Retinoids, edited by M. B. Sporn, A. B. Roberts, and D. S.Goodman. New York:Raven.1994,387-441.
[8]Ross, S.A., De Luca, L.M. et al. Retinoids in embryonal development. Physiol. Rev.2000,80: 1021-1054.
[9]Kochhar, D.M., Johnson, E.M. Morphological and autoradiographic studies of cleft palate induced in rat embryos by maternal hypervitaminosis A. J. Embryol. Exp. Morphol.1965,14: 223-238.
[10]Abbott, B.D., Harris, M.W., Birnbaum, L.S. Etiology of retinoic acidinduced cleft palate varies with the embryonic stage. Teratology.1989,40:533-553.
[11]Kochhar, D.M., Johnson. E.M. Morphological and autoradiographic studies of cleft palate induced in rat embryos by maternal hypervitaminosis A. J. Embryol. Exp. Morphol.1965,14: 223-238.
[12]Giroud, A.& Martinet, M. Teratogenese par hautes doses de vitamine A en fonction des stades du developpement. Archs. Anat. Microsc.1956,45:77-98.
[13]Junko Okano, Shigehiko Suzuki, Kohei Shiota. Involvement of apoptotic cell death and cell cycle perturbation in retinoicacid-induced cleft palate in mice. Toxicology and Applied Pharmacology.2007,221:42-56.
[14]Padmanabhan, R. and A. Ijaz. Retinoic acid-induced asymmetric craniofacial growth and cleft palate in the to mouse fetus. Reproductive Toxicology.1997,11(6):843-860.
[15]Okano, J., S. Suzuki, et al. Involvement of apoptotic cell death and cell cycle perturbation in retinoic acid-induced cleft palate in mice. Toxicology and Applied Pharmacology.2007,221(1): 42-56.
[16]Tseng, C. F., C. C. Lin, et al. Antioxidant role of human haptoglobin. PROTEOMICS.2004, 4 (8):2221-2228.
[17]Saso, L., P. Tommasino, et al. Changes of acute-phase proteins in streptozotocin-induced diabetic rats. Physiol Res.2000,49(4):403-9.
[18]Van Vlierberghe, H., M. Langlois, et al. Haptoglobin polymorphisms and iron homeostasis in health and in disease. Clinica Chimica Acta.2004,345(1-2):35-42.
[19]Bicho, M. C., A. Pereira da Silva, et al. Sex steroid hormones influence the risk for cervical cancer:modulation by haptoglobin genetic polymorphism. Cancer Genetics and Cytogenetics. 2000,191(2):85-89.
[20]Hamrita, B., K. Chahed, et al. Proteomics-based identification of [alpha]1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas. Clinica Chimica Acta.2009,404(2):111-118.
[21]Vanhooren, V., X.-E. Liu, et al. N-glycan profiles as tools in diagnosis of hepatocellular carcinoma and prediction of healthy human ageing. Mechanisms of Ageing and Development. 2008,130(1-2):92-97.
[22]Istvan Liko, Marianna Mak, et al. Evidence for an extended interacting surface between [beta]-amyloid and serum amyloid P component. Neuroscience Letters.2007,412(1):51-55.
[23]Song, Z., L. Cai, et al. Accumulation and expression of serum amyloid P component in human atherosclerotic lesions. Atherosclerosis In Press, Corrected Proof.
[24]Szilvia Veszelka, Zoltan Urbanyi, et al. Human serum amyloid P component attenuates the bacterial lipopolysaccharide-induced increase in blood-brain barrier permeability in mice. Neuroscience Letters.2003,352(1):57-60.
[25]Urbanyi, Z., Lajos Laszlo, et al. Serum amyloid P component induces neuronal apoptosis and [beta]-amyloid immunoreactivity. Brain Research.2003,988(1-2):69-77.
[26]Zoltan Urbanyi, Miklos Sass, et al.Serum amyloid P component induces TUNEL-positive nuclei in rat brain after intrahippocampal administration. Brain Research.2007,1145:221-226.
[27]Vamsidhara C Dhulipala. Cell cycle proteins in normal and chemicallyinduced abnormal secondary palatedevelopment:a review [J]. Human & Experimental Toxicology (2006) 25:675-682.
[28]李鑫,金岩等.细胞程序性死亡及抗凋亡基因bcl-2在C57BL6N系小鼠腭裂形成中的意义[J]现代预防医学.2007,34(5)：858-860.
[29]李杨,石冰.非综合征唇腭裂的候选基因位点.口腔颌面外科杂志.2007,17(1)：82-85
[30]王文恭,童坦君.细胞凋亡过程中bacp-2基因的甲基化。[J]中国生物化学与分子生物学报。1998,14(3)：309-313.
[31]王海燕,王启会.细胞凋亡的相关基因调控.现代预防医学.2007,34(5)858-860.王如,刘彬.腭发育不同时期全反式视黄酸对腭突细胞增殖和凋亡的影响.华西口腔医学杂志.2008,26(5)：546-549.
[32]李鑫,金岩.细胞凋亡及p53基因的表达在C57BL6N系小鼠腭裂形成中的意义.实用口腔医学杂志.2000,16(6)：440-442.
[33]Jiu-Zhen Jin and Jixiang Ding. Analysis of cell migration, transdifferentiation and apoptosisduring mouse secondary palate fusion. Development 2006,133:3341-3347
[34]Gregg Duester. Retinoic Acid Regulation of the Somitogenesis Clock. Birth Defects Research 2007,81:84-92
[35]Vamsidhara C Dhulipalal, Wade V Welshons. Cell cycle proteins in normal and chemically induced abnormal secondary palatedevelopment:a revieW Human & Experimental Toxicology 2006,25:675-68
[1]Aivaliotis, M., et al., Large-scale identification of N-terminal peptides in the halophilic archaea Halobacterium salinarum and Natronomonas pharaonis. Journal of Proteome Research,2007.6(6): p.2195-2204.
[2]Kuyama, H., et al., A method for N-terminal de novo sequence analysis of proteins by matrix-assisted laser desorption/ionization mass spectrometry. Analytical Biochemistry,2008. 380(2):p.291-296.
[3]Roemer, I., et al., Analysis of mouse beta-haptoglobin chain by lectin affinoblotting detection. Electrophoresis,2001.22(14):p.3038-3042.
[4]Golaz, O., et al., Rapid Detection of the Main Human Plasma Glycoproteins by 2-Dimensional Polyacrylamide-Gel Electrophoresis Lectin Affinoblotting. Electrophoresis,1995.16(7):p. 1187-1189.
[5]Marynen, P., et al., A Genetic-Polymorphism in a Functional Domain of Human-Pregnancy Zone Protein-the Bait Region-Genomic Structure of the Bait Domains of Human-Pregnancy Zone Protein and Alpha-2 Macroglobulin. Febs Letters,1990.262(2):p.349-352.
[6]Claesson, R., et al., A new cleavage site for elastase within the complement component 3. Apmis,2010.118(10):p.765-768.
[7]Horie, M., et al., The N-terminal region is important for the nuclease activity and thermostahility of the flap endonuclease-1 from Sulfolobus tokodaii. Bioscience Biotechnology and Biochemistry,2007.71(4):p.855-865.
[8]Huang, Z.H., et al., Protein sequencing by matrix-assisted laser desorption ionization-postsource decay mass spectrometry analysis of the N-tris(2,4,6-trimethoxyphenyl)phosphine-acetylated tryptic digests. Analytical Biochemistry, 1999.268(2):p.305-317.
[9]An, M.R., et al., Efficient and clean charge derivatization of peptides for analysis by mass spectrometry. Rapid Communications in Mass Spectrometry,2010.24(13):p.1869-1874.
[10]Chamot-Rooke, J., et al., The combination of electron capture dissociation and fixed charge derivatization increases sequence coverage for O-glycosylated and O-phosphorylated peptides. Journal of the American Society for Mass Spectrometry,2007.18(8):p.1405-1413.
[11]Gallien, S., et al., Ortho-proteogenomics:Multiple proteomes investigation through orthology and a new MS-based protocol. Genome Research,2009.19(1):p.128-135.
[12]Franck, J., et al., On-Tissue N-Terminal Peptide Derivatizations for Enhancing Protein Identification in MALDI Mass Spectrometric Imaging Strategies. Analytical Chemistry,2009. 81(20):p.8305-8317.
[l]钱小红,贺福初.蛋白质组学：理论与方法.北京：科学出版社,2003.
[2]夏其昌,曾嵘.蛋白质化学与蛋白质组学.北京：科学出版社,2004.
[3]Wasinger, V. C., Cordwell, S. J., Cerpa-Poljak, A., Yan, J. X., Gooley, A. A., Wilkins, M. R., Duncan, M. W., Harris, R., Williams, K. L., and Humphery-Smith, I. (1995). Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis 16,1090-4.
[4]Klose, J. (1975). Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals. Humangenetik 26,231-43.
[5]O'Farrell, P. H. (1975). High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250,4007-21.
[6]Rabilloud, T., Chevallet, M., Luche, S., and Lelong, C. Two-dimensional gel electrophoresis in proteomics:Past, present and future. Journal of Proteomics 73,2064-2077.
[7]Karas, M., and Hillenkamp, F. (1988). Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. Anal Chem 60,2299-301.
[8]Gygi, S. P., Rist, B., Gerber, S. A., Turecek, F., Gelb, M. H., and Aebersold, R. (1999). Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat Biotechnol 17,994-9.
[9]Gygi, S. P., Rist, B., Griffin, T. J., Eng, J., and Aebersold, R. (2002). Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags. J Proteome Res 1,47-54.
[10]Tseng, C. F., Lin, C. C., Huang, H. Y., Liu, H. C., and Mao, S. J. T. (2004). Antioxidant role of human haptoglobin. PROTEOMICS 4,2221-2228.