不同温度对虎奶菇胞内胞外蛋白差异表达的影响
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摘要
自溶现象广泛存在于各种微生物中,但有关食药用菌虎奶菇(Pleurotustuber-regium(Fr.)Sing)菌丝体自溶的研究国内外均未见报道。本文通过筛选获得引起自溶的较佳条件,用该条件进行虎奶菇自溶处理,最终应用差异蛋白质组分析方法研究其自溶。
首先利用液体发酵法培养虎奶菇菌丝体,分别研究了不同温度、pH值与渗透压等因素对其菌丝体自溶的影响。结果表明,虎奶菇菌丝体较佳的单因素自溶条件分别为温度40℃、pH 9.0、NaCl浓度3-4%。
然后比较了虎奶菇菌丝体蛋白质的三种提取方法以寻找一种适用于蛋白质组学双向电泳的高分辨率的提取方法。结果表明TCA/丙酮法提取所得到的蛋白质样品的双向电泳图谱效果最佳。同时获得制备胞外蛋白样品的方法。
最终以40℃处理为实验组,10℃处理为对照组,采用差异蛋白质组学方法研究虎奶菇的自溶现象,得到22个明显上调的蛋白质点,经MALDI-TOF/MS检测及肽指纹图谱(PMF)分析,其中8个蛋白质得到鉴定,表现为:蛋白酶体参与泛素-蛋白酶体通路,β-1,3-glucanosyltransferase及微管蛋白与细胞壁的更新、细胞壁的修补相关,蛋白酶和内切葡聚糖酶促进细胞壁和细胞质自然降解,14-3-3蛋白参与细胞信号转导。因此推测菌丝体自溶过程如下:当营养条件缺乏或高温下,往往出现细胞正常代谢失控,细胞内部结构解体,这时蛋白酶体承担了降解细胞内蛋白质的任务,蛋白酶和内切葡聚糖酶等水解酶的大量释放促进细胞壁和细胞质降解,获得的自溶产物为其后代生长提供营养源;大量合成β-1,3-glucanosyltransferase及微管蛋白等,参与细胞壁的更新、修补,以维持其正常形态和功能。
Autolysis phenomena exist widely in a variety of microbes.However,the mycelium autolysis of the edible and medical Pleurotus tuber-regium(Fr.) has not been reported at home and abroad.This experiment initiated from screening the appropriate conditions of Pleurotus tuber-regium(Fr.) mycelium autolysis,then treating the mycelium with the optimal autolysis condition,and finally analyzing the autolysis by using differential proteomic method.
First,the effects of different temperature,pH and osmotic pressure on mycelium autolysis were studied via liquid fermentation of Pleurotus tuber-regium(Fr.) mycelium.The results showed that the appropriate monofactorial condition of autolysis is temperature 40℃,pH 9.0 and NaCl concentration 3-4%.
Then,an efficient method for extracting the total proteins from the Pleurotus tuber-regium(Fr.) mycelium was established with high resolution applicable to the proteomic 2-D electrophoresis analysis.The results indicated that the TCA/acetone method was optimal for extracting total protein for 2-D electrophoresis.The method of extracting extracellular protein was also obtained in this research.
Lastly,the autolysis of Pleurotus tuber-regium(Fr.) mycelium was investigated by differential proteomics,using 40℃treatment as experimental group and 10℃as control.Twenty-two protein spots with remarkable up-regulation were obtained for analysis by MALDI-TOF/MS and PMF,and eight proteins were identified.Of these proteins,proteasome participates in ubiquitin-proteasome pathway;β-1,3-glucanosyltransferase and tubulin are involved in the renew and repair of cell wall;protease and endo-glucanase promote the natural degradation of cell wall and cytoplasm;and 14-3-3 protein deals with cell signal transduction.Thus,the process of Pleurotus tuber-regium(Fr.) mycelium autolysis is presumed as follows:under nutrition shortage or high temperature treatment,the cells undergo metabolic malfunction and inner structure disruption;then the proteasome degrades the intracellular proteins and leads to the release of hydrolases such as prolease and endo-glucanase to accelerate cell wall and cytoplasm degradation;the autolysis products serve as the nutrition for the progeny;and theβ-1,3-glucanosyltransferase and tubulin maintain the normal cell shape and function by renewing and repairing cell wall.
首先利用液体发酵法培养虎奶菇菌丝体,分别研究了不同温度、pH值与渗透压等因素对其菌丝体自溶的影响。结果表明,虎奶菇菌丝体较佳的单因素自溶条件分别为温度40℃、pH 9.0、NaCl浓度3-4%。
然后比较了虎奶菇菌丝体蛋白质的三种提取方法以寻找一种适用于蛋白质组学双向电泳的高分辨率的提取方法。结果表明TCA/丙酮法提取所得到的蛋白质样品的双向电泳图谱效果最佳。同时获得制备胞外蛋白样品的方法。
最终以40℃处理为实验组,10℃处理为对照组,采用差异蛋白质组学方法研究虎奶菇的自溶现象,得到22个明显上调的蛋白质点,经MALDI-TOF/MS检测及肽指纹图谱(PMF)分析,其中8个蛋白质得到鉴定,表现为:蛋白酶体参与泛素-蛋白酶体通路,β-1,3-glucanosyltransferase及微管蛋白与细胞壁的更新、细胞壁的修补相关,蛋白酶和内切葡聚糖酶促进细胞壁和细胞质自然降解,14-3-3蛋白参与细胞信号转导。因此推测菌丝体自溶过程如下:当营养条件缺乏或高温下,往往出现细胞正常代谢失控,细胞内部结构解体,这时蛋白酶体承担了降解细胞内蛋白质的任务,蛋白酶和内切葡聚糖酶等水解酶的大量释放促进细胞壁和细胞质降解,获得的自溶产物为其后代生长提供营养源;大量合成β-1,3-glucanosyltransferase及微管蛋白等,参与细胞壁的更新、修补,以维持其正常形态和功能。
Autolysis phenomena exist widely in a variety of microbes.However,the mycelium autolysis of the edible and medical Pleurotus tuber-regium(Fr.) has not been reported at home and abroad.This experiment initiated from screening the appropriate conditions of Pleurotus tuber-regium(Fr.) mycelium autolysis,then treating the mycelium with the optimal autolysis condition,and finally analyzing the autolysis by using differential proteomic method.
First,the effects of different temperature,pH and osmotic pressure on mycelium autolysis were studied via liquid fermentation of Pleurotus tuber-regium(Fr.) mycelium.The results showed that the appropriate monofactorial condition of autolysis is temperature 40℃,pH 9.0 and NaCl concentration 3-4%.
Then,an efficient method for extracting the total proteins from the Pleurotus tuber-regium(Fr.) mycelium was established with high resolution applicable to the proteomic 2-D electrophoresis analysis.The results indicated that the TCA/acetone method was optimal for extracting total protein for 2-D electrophoresis.The method of extracting extracellular protein was also obtained in this research.
Lastly,the autolysis of Pleurotus tuber-regium(Fr.) mycelium was investigated by differential proteomics,using 40℃treatment as experimental group and 10℃as control.Twenty-two protein spots with remarkable up-regulation were obtained for analysis by MALDI-TOF/MS and PMF,and eight proteins were identified.Of these proteins,proteasome participates in ubiquitin-proteasome pathway;β-1,3-glucanosyltransferase and tubulin are involved in the renew and repair of cell wall;protease and endo-glucanase promote the natural degradation of cell wall and cytoplasm;and 14-3-3 protein deals with cell signal transduction.Thus,the process of Pleurotus tuber-regium(Fr.) mycelium autolysis is presumed as follows:under nutrition shortage or high temperature treatment,the cells undergo metabolic malfunction and inner structure disruption;then the proteasome degrades the intracellular proteins and leads to the release of hydrolases such as prolease and endo-glucanase to accelerate cell wall and cytoplasm degradation;the autolysis products serve as the nutrition for the progeny;and theβ-1,3-glucanosyltransferase and tubulin maintain the normal cell shape and function by renewing and repairing cell wall.
引文
1.包水明,李荣同,陈传红等.虎奶菇及其人工栽培技术[J].食用菌,2007,29(3):63-64.
2.吴锦忠.虎奶菇和虎乳灵芝液体发酵及化学成分分析[D].福建农林大学博士学位论文,2005,4.
3.Fuensanta Reyes,R.Lahoz,M.J.Martinez,C.Alfonso.Chitosanases in the autolysis of Mucor rouxii[J].Mycopathologia,1985,89:181-187.
4.田永强.微生物自溶现象研究进展[J].微生物学杂志.1997,17(2):52-60.
5.M.I.Perez-Leblic,F.Reyes,M.J.Martinez,R.Lahoz.Cell wall degradation in the auolysis of filamentons fungi[J].Mycopathologia,1982,80:147-155.
6.宁正祥.酵母自溶动力学研究[J].微生物学报,1994,34(3):213-219.
7.柔刚,许瑶.啤酒酵母的自溶现象极其防治措施[J].酿酒科技,1998,6:21-22.
8.Lahoz R,Reyes F,Perez Lebli MI.lyric enzymes in the autolysis of fungi[J].Mycopathologia,1976,60(1):45-48.
9.李祥,彭莉,王毅,等.酵母自溶研究[J].中国酿造,2001,(4):17-19.
10.吴宝平,陶树兴.酵母细胞最适生长条件和自溶条件研究[J].陕西师范大学学报(自然科学版),2004,32:81-85.
11.孙向军,姚晓敏,张伟.酵母细胞自溶条件的研究[J],上海农学院学报,2000,18(1):36-39.
12.高玉荣.啤酒废酵母自溶条件的研究.酿酒科技[J],2002(2):74-76.
13.方尚玲,马丽,吴思方.影响酵母自溶因素与条件的研究[J].粮食与饲料工业,2000,3:21-23.
14.张晓鸣,袁信华,章克昌.啤酒废酵母自溶动力学的研究[J].无锡轻工大学学报,2000,19(5):440-442.
15.曾凡坤,汤务霞.提高酵母抽提物得率的工艺条件[J].食品与发酵工业,2004,30(8):25-29.
16.王校红.功能性食品添加剂一酵母自溶物的生产特性和用途[J].黑龙江粮油科技,2001,(2):57-58.
17.钟文晖.利用啤酒废酵母研制营养调味品[J].食品科学,1996,(2):15-21.
18.田卓玲,于慧敏,沈忠耀.用于微生物培养基有机氮源的废酵母自溶液研究[J].食品与发酵工业,2003,29(5):1-6.
19.候春,丁骅孙.米曲霉自溶作用的研究Ⅰ:米曲霉的自溶条件[J].云南大学学报,1995,17(3):275-277.
20.候春,丁骅孙.米曲霉自溶作用的研究II:自溶过程中细胞物质及酶活的变化[J].云南大学学报,1996,18(2):146-148.
21.贺荣平.食用菌系列休闲食品的开发与研究[J].加工技术,2005,(3):37-40.
22.邵伟,刘世玲,熊泽,等.保健香菇醋的研制[J].食品工业,2001,(5):27-28.
23.黄年来,郭美英,黄黎红.虎奶菇及其栽培[J].中国食用菌,1997,16(1):13-13.
24.江枝和,李三署,郑永标,等.虎奶菇菌核和子实体微量元素及氨基酸分析[J].食用菌学报,2000,7(2):47-50.
25.吴锦忠,张志强,张丽明,等.虎奶菇化学成分分析[J].福建中医学院学报,2004,14(1):8-10.
26.夏其昌,曾嵘等.蛋白质化学与蛋白质组学[M].科学出版社,2005.
27.李林,吴家睿,李伯良.蛋白质组学的产生及其重要意义[J].生命科学,1999,11(2):49-50.
28.龙晓辉,莫志宏,张耀洲.基于二维凝胶电泳的蛋白质定量分析技术[J].化学进展,2006,18(4):474-481.
29.袁雪宇.差异蛋白质组学技术和应用前景[J].同济大学学报(医学版),2004,25(4):349-351.
30.罗治文.质谱技术研究进展[J].国外医学生物医学工程分册,2005,28(3):134-137.
31.R.Graham Cooks,Zheng Ouyang,Zoltan Takats,Justin M.Wiseman.Ambient Mass Spectrometry[J].science,2006,311:1566-1570.
32.吴敏曼,孙虹.蛋白质组学及在鼻黏膜炎症性疾病研究中应用[J].国外医学耳鼻咽喉科学分册,2005,29(5):298-301.
33.费嘉,马文丽,郑文岭.蛋白质芯片在蛋白质组研究中的作用[J].生命科学,2005,17(2):132-136.
34.焦立新.烟曲霉致病机制的蛋白质组学研究[D].吉林大学博士学位论文,2007,4.
35.Hodges PE,Mckeee AHZ,Davis B RThe yeast proteome database(YPD):a model for the organization and presentation of genome-wide functional data[J].Nucleic Acids Research,1999,27:69-73.
36.AKumar.et al.Subcellular localization of the yeast proteome[J].Genes &development,2002,16:707-719.
37.http://www.ebiotrade.com/newsf/2006-1/2006123153232.htm
38.Steffen Ohlmeier.et al.The Yeast Mitochondrial Proteome,a Study of Fermentative and Respiratory Growth[J].Biol.Chem.,February 6,2004,279(6):3956-3979.
39.廖国建,陈洪斌等.应用蛋白质组学技术研究铬(Ⅵ)对粟酒裂殖酵母的分子毒性影响[J].环境科学学报,2005,25(12):1693-1697.
40.EMelin,J.Schnurer,E.G.H.Wagner.Proteome analysis of Aspergillus nidulans reveals proteins associated with the response to the antibiotic concanamycin A,produced by Streptomyces species[J].Mol Genet Genomics,2002,267:695-702.
41.Bruneau.et al.Proteome analysis of Aspergillus fumigatus identifies glycosylphosphatidylinositol-anchored proteins associated to the cell wall biosynthesis[J].Electrophoresis,2002,22:2812-2823.
42.Olaf Kniemeyer.et al.Optimisation of a 2-D gel electrophoresis protocol for the human-pathogenic fungus Aspergillus fumigatus[J].Curr Genet,2006,49:178-189.
43.Ambrosino P.et al.extracellular proteome of trichoderma harzianum to identify proteins with biotechnological value[J].Joural of zhejiang university(Agric.&Life Sci),2004,30(4):452.
44.M.Belen Suarez et al.Proteomic analysis of secreted proteins from Trichoderma harzianum Identification of a fungal cell wall-induced aspartic protease[J].Fungal Genetics and Biology,2005,42:924-934.
45.Amber Vanden Wymelenberg.et al.Computational analysis of the Phanerochaete chrysosporium v2.0 genome database and mass spectrometry identification of peptides in ligninolytic cultures reveal complex mixtures of secreted proteins[J].Fungal Genetics and Biology,2006,1087-1845.
46.胡开辉.微生物学实验[M].中国林业出版社(北京),2004:215-215.
47.汪家政,范明主编.蛋白质技术手册[M].科学出版社(北京),2000:42-46.
48.张国林,王三英.采用比较蛋白质组学方法研究p38激酶对239T细胞蛋白质表达的影响[J].厦门大学学报(自然科学版),2005,44:123-127.
49.陈茂营,徐文方.蛋白酶体与蛋白酶体抑制剂[J].中国药物化学杂志.2007,17(4):249-253.
50.金伟军,姚祥春,吕美巧等.泛素-蛋白酶体系统的结构、作用和调控机制[J].科技通报,2008,24(1):29-34.
51.Jayhyuk Myung,Kyung Bo Kim,and Craig M.Crews.The Ubiquitin-Proteasome Pathway and Proteasome Inhibitors[J].Med Res Rev.2001,21(4):245-273.
52.赵宝昌,崔秀云.蛋白酶体的结构与功能[J].生命的化学.2002,22(3):215-218.
53.Coux O,Tanaka K,Goldberg AL.Structure and functions of the 20S and 26S proteasomes[J].Ann Rev Biochem,1996,65:801-847.
54.Elena Koulich,Xiaohua Li,and George N.DeMartino.Relative Structural and Functional Roles of Multiple Deubiquitylating Proteins Associated with ammalian 26S Proteasome[J].Molecular Biology of the Cell,2008,19,1072-1082.
55.Tanaka K.Proteasomes:structure and biology[J].J Biochem(tokyo),1998,123:195-204.
56.陈国强,张海婧,李智立.蛋白质组学技术在泛素.蛋白酶体系统研究中的应用[J].基础医学与临床,2007,27(1):1-7.
57.董辉,李越中,胡玮.γ-微管蛋白研究进展[J].生物化学与生物物理进展,2002,29(5):686-690.
58.时兰春,王益川,王伯初.植物细胞骨架与细胞生长[J].植物生理学通讯.2007,43(6):1175-1181.
59.陈晓燕,汪志平,杨灵勇.原核细胞骨架蛋白的结构与功能[J].细胞生物学杂志,2006,28:699-703.
60.许子勤,李守全,孙敬三.微管与微管蛋白概述及其研究进展[J].细胞生物学杂志,1989,11(2):54-58.
61.Robbert RHartland.et al.A Novel β-(1,3)-glucanosyltransferase from the Cell Wall of Aspergillus fumigatus[J].BIOLOGICAL CHEMISTRY.1996,271(43):26843-26849.
62.http://baike.baidu.com/view/782157.htm
63.欧阳杰,蒋建雄,张天真,郭旺珍,易自力.内切-1,4-β-葡聚糖酶在植物细胞生长发育中的作用[J].西北植物学报,2007,4:206-213.
64.Pierre Beguin.Molecular biology of cellulose degradation[D].Microbiol.1990,44:219-248.
65.伍家发,吴乔.14-3-3蛋白家族的调控机制和生物学功能[J].细胞生物学杂志,2005.27:101-104.
66.Hrvoje Fulgosi,Jǖrgen Soll,Simone de Faria Maraschin,Henrie A.A.J.Korthout,Mei Wang and Christa Testerink.14-3-3 proteins and plant development[J].Plant Molecular Biology,2002,50(6):1019-1029.
67.文彬,王小菁.14-3-3蛋白研究进展[J].生命科学,2004,16(4):226-230.
2.吴锦忠.虎奶菇和虎乳灵芝液体发酵及化学成分分析[D].福建农林大学博士学位论文,2005,4.
3.Fuensanta Reyes,R.Lahoz,M.J.Martinez,C.Alfonso.Chitosanases in the autolysis of Mucor rouxii[J].Mycopathologia,1985,89:181-187.
4.田永强.微生物自溶现象研究进展[J].微生物学杂志.1997,17(2):52-60.
5.M.I.Perez-Leblic,F.Reyes,M.J.Martinez,R.Lahoz.Cell wall degradation in the auolysis of filamentons fungi[J].Mycopathologia,1982,80:147-155.
6.宁正祥.酵母自溶动力学研究[J].微生物学报,1994,34(3):213-219.
7.柔刚,许瑶.啤酒酵母的自溶现象极其防治措施[J].酿酒科技,1998,6:21-22.
8.Lahoz R,Reyes F,Perez Lebli MI.lyric enzymes in the autolysis of fungi[J].Mycopathologia,1976,60(1):45-48.
9.李祥,彭莉,王毅,等.酵母自溶研究[J].中国酿造,2001,(4):17-19.
10.吴宝平,陶树兴.酵母细胞最适生长条件和自溶条件研究[J].陕西师范大学学报(自然科学版),2004,32:81-85.
11.孙向军,姚晓敏,张伟.酵母细胞自溶条件的研究[J],上海农学院学报,2000,18(1):36-39.
12.高玉荣.啤酒废酵母自溶条件的研究.酿酒科技[J],2002(2):74-76.
13.方尚玲,马丽,吴思方.影响酵母自溶因素与条件的研究[J].粮食与饲料工业,2000,3:21-23.
14.张晓鸣,袁信华,章克昌.啤酒废酵母自溶动力学的研究[J].无锡轻工大学学报,2000,19(5):440-442.
15.曾凡坤,汤务霞.提高酵母抽提物得率的工艺条件[J].食品与发酵工业,2004,30(8):25-29.
16.王校红.功能性食品添加剂一酵母自溶物的生产特性和用途[J].黑龙江粮油科技,2001,(2):57-58.
17.钟文晖.利用啤酒废酵母研制营养调味品[J].食品科学,1996,(2):15-21.
18.田卓玲,于慧敏,沈忠耀.用于微生物培养基有机氮源的废酵母自溶液研究[J].食品与发酵工业,2003,29(5):1-6.
19.候春,丁骅孙.米曲霉自溶作用的研究Ⅰ:米曲霉的自溶条件[J].云南大学学报,1995,17(3):275-277.
20.候春,丁骅孙.米曲霉自溶作用的研究II:自溶过程中细胞物质及酶活的变化[J].云南大学学报,1996,18(2):146-148.
21.贺荣平.食用菌系列休闲食品的开发与研究[J].加工技术,2005,(3):37-40.
22.邵伟,刘世玲,熊泽,等.保健香菇醋的研制[J].食品工业,2001,(5):27-28.
23.黄年来,郭美英,黄黎红.虎奶菇及其栽培[J].中国食用菌,1997,16(1):13-13.
24.江枝和,李三署,郑永标,等.虎奶菇菌核和子实体微量元素及氨基酸分析[J].食用菌学报,2000,7(2):47-50.
25.吴锦忠,张志强,张丽明,等.虎奶菇化学成分分析[J].福建中医学院学报,2004,14(1):8-10.
26.夏其昌,曾嵘等.蛋白质化学与蛋白质组学[M].科学出版社,2005.
27.李林,吴家睿,李伯良.蛋白质组学的产生及其重要意义[J].生命科学,1999,11(2):49-50.
28.龙晓辉,莫志宏,张耀洲.基于二维凝胶电泳的蛋白质定量分析技术[J].化学进展,2006,18(4):474-481.
29.袁雪宇.差异蛋白质组学技术和应用前景[J].同济大学学报(医学版),2004,25(4):349-351.
30.罗治文.质谱技术研究进展[J].国外医学生物医学工程分册,2005,28(3):134-137.
31.R.Graham Cooks,Zheng Ouyang,Zoltan Takats,Justin M.Wiseman.Ambient Mass Spectrometry[J].science,2006,311:1566-1570.
32.吴敏曼,孙虹.蛋白质组学及在鼻黏膜炎症性疾病研究中应用[J].国外医学耳鼻咽喉科学分册,2005,29(5):298-301.
33.费嘉,马文丽,郑文岭.蛋白质芯片在蛋白质组研究中的作用[J].生命科学,2005,17(2):132-136.
34.焦立新.烟曲霉致病机制的蛋白质组学研究[D].吉林大学博士学位论文,2007,4.
35.Hodges PE,Mckeee AHZ,Davis B RThe yeast proteome database(YPD):a model for the organization and presentation of genome-wide functional data[J].Nucleic Acids Research,1999,27:69-73.
36.AKumar.et al.Subcellular localization of the yeast proteome[J].Genes &development,2002,16:707-719.
37.http://www.ebiotrade.com/newsf/2006-1/2006123153232.htm
38.Steffen Ohlmeier.et al.The Yeast Mitochondrial Proteome,a Study of Fermentative and Respiratory Growth[J].Biol.Chem.,February 6,2004,279(6):3956-3979.
39.廖国建,陈洪斌等.应用蛋白质组学技术研究铬(Ⅵ)对粟酒裂殖酵母的分子毒性影响[J].环境科学学报,2005,25(12):1693-1697.
40.EMelin,J.Schnurer,E.G.H.Wagner.Proteome analysis of Aspergillus nidulans reveals proteins associated with the response to the antibiotic concanamycin A,produced by Streptomyces species[J].Mol Genet Genomics,2002,267:695-702.
41.Bruneau.et al.Proteome analysis of Aspergillus fumigatus identifies glycosylphosphatidylinositol-anchored proteins associated to the cell wall biosynthesis[J].Electrophoresis,2002,22:2812-2823.
42.Olaf Kniemeyer.et al.Optimisation of a 2-D gel electrophoresis protocol for the human-pathogenic fungus Aspergillus fumigatus[J].Curr Genet,2006,49:178-189.
43.Ambrosino P.et al.extracellular proteome of trichoderma harzianum to identify proteins with biotechnological value[J].Joural of zhejiang university(Agric.&Life Sci),2004,30(4):452.
44.M.Belen Suarez et al.Proteomic analysis of secreted proteins from Trichoderma harzianum Identification of a fungal cell wall-induced aspartic protease[J].Fungal Genetics and Biology,2005,42:924-934.
45.Amber Vanden Wymelenberg.et al.Computational analysis of the Phanerochaete chrysosporium v2.0 genome database and mass spectrometry identification of peptides in ligninolytic cultures reveal complex mixtures of secreted proteins[J].Fungal Genetics and Biology,2006,1087-1845.
46.胡开辉.微生物学实验[M].中国林业出版社(北京),2004:215-215.
47.汪家政,范明主编.蛋白质技术手册[M].科学出版社(北京),2000:42-46.
48.张国林,王三英.采用比较蛋白质组学方法研究p38激酶对239T细胞蛋白质表达的影响[J].厦门大学学报(自然科学版),2005,44:123-127.
49.陈茂营,徐文方.蛋白酶体与蛋白酶体抑制剂[J].中国药物化学杂志.2007,17(4):249-253.
50.金伟军,姚祥春,吕美巧等.泛素-蛋白酶体系统的结构、作用和调控机制[J].科技通报,2008,24(1):29-34.
51.Jayhyuk Myung,Kyung Bo Kim,and Craig M.Crews.The Ubiquitin-Proteasome Pathway and Proteasome Inhibitors[J].Med Res Rev.2001,21(4):245-273.
52.赵宝昌,崔秀云.蛋白酶体的结构与功能[J].生命的化学.2002,22(3):215-218.
53.Coux O,Tanaka K,Goldberg AL.Structure and functions of the 20S and 26S proteasomes[J].Ann Rev Biochem,1996,65:801-847.
54.Elena Koulich,Xiaohua Li,and George N.DeMartino.Relative Structural and Functional Roles of Multiple Deubiquitylating Proteins Associated with ammalian 26S Proteasome[J].Molecular Biology of the Cell,2008,19,1072-1082.
55.Tanaka K.Proteasomes:structure and biology[J].J Biochem(tokyo),1998,123:195-204.
56.陈国强,张海婧,李智立.蛋白质组学技术在泛素.蛋白酶体系统研究中的应用[J].基础医学与临床,2007,27(1):1-7.
57.董辉,李越中,胡玮.γ-微管蛋白研究进展[J].生物化学与生物物理进展,2002,29(5):686-690.
58.时兰春,王益川,王伯初.植物细胞骨架与细胞生长[J].植物生理学通讯.2007,43(6):1175-1181.
59.陈晓燕,汪志平,杨灵勇.原核细胞骨架蛋白的结构与功能[J].细胞生物学杂志,2006,28:699-703.
60.许子勤,李守全,孙敬三.微管与微管蛋白概述及其研究进展[J].细胞生物学杂志,1989,11(2):54-58.
61.Robbert RHartland.et al.A Novel β-(1,3)-glucanosyltransferase from the Cell Wall of Aspergillus fumigatus[J].BIOLOGICAL CHEMISTRY.1996,271(43):26843-26849.
62.http://baike.baidu.com/view/782157.htm
63.欧阳杰,蒋建雄,张天真,郭旺珍,易自力.内切-1,4-β-葡聚糖酶在植物细胞生长发育中的作用[J].西北植物学报,2007,4:206-213.
64.Pierre Beguin.Molecular biology of cellulose degradation[D].Microbiol.1990,44:219-248.
65.伍家发,吴乔.14-3-3蛋白家族的调控机制和生物学功能[J].细胞生物学杂志,2005.27:101-104.
66.Hrvoje Fulgosi,Jǖrgen Soll,Simone de Faria Maraschin,Henrie A.A.J.Korthout,Mei Wang and Christa Testerink.14-3-3 proteins and plant development[J].Plant Molecular Biology,2002,50(6):1019-1029.
67.文彬,王小菁.14-3-3蛋白研究进展[J].生命科学,2004,16(4):226-230.