随着“蛋白质组学”这一概念的出现和发展,细胞分子网络领域的研究取得了巨大的进展,人们对此领域的认识也日益深入。然而,蛋白质的冗余性、动力学特点和互作作用,使相关研究面临巨大挑战。在本书中,多位专家阐述了蛋白质组学的各种研究技术,纵览了本领域各研究方向的难点与可能性,提供了最新的实验方案和具体实例。作为《分子生物学方法》系列丛书的一卷,本书简明易懂,各章均包含针对标题的导言、必备材料与试剂的清单、分步骤且易于操作的实验室方案、疑难问题的注意事项和易犯失误的避免。本书专业权威、易于使用,适合作为实验室指南用书,可以激发读者对蛋白质组学这一复杂且重要的领域的研究兴趣。
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本书专业权威、易于使用,适合作为实验室指南用书,可以激发读者对蛋白质组学这一复杂且重要的领域的研究兴趣。
目录
前言 v
撰稿人 ix
第一部分 引言
1. 蛋白质组学简介 3
Friedrich Lottspeich
第二部分 电泳分离
2. 高分辨率二维电泳 13
Walter Weiss and Angelika Gorg
3. 非经典二维电泳 33
JacquelineBurre,Ilka Wittig,and Hermann Schagger
4. 蛋白质检测与定量技术用于基于凝胶的蛋白质组学分析 59
Walter Weiss,Florian Weiland,and Angelika Gorg
第三部分 质谱和串联质谱的应用
5. 基质辅助激光解吸电离质谱 85
Rainer Cramer
6. 毛细管电泳质谱联用应用于人尿液蛋白质组学分析与生物标志物发现 105
PetraZurbig,Eric Schiffer,and Harald Mischak
7. 多肽纳升级液相色谱指南 123
Thomas Frohlich and Georg J.Arnold
8. 多维蛋白鉴定技术 143
Katharina Lohrig and Dirk Wolters
9. 肽段为中心的蛋白质组学技术分析血小板蛋白 155
Oliver Simon,Stefanie Wortelkamp,and Albert Sickmann
10. 高分辨率质谱蛋白质组分析技术鉴定小鼠小肠20S蛋白质组分子组成 173
Reinhold Weber,Regina Preywisch,Nikolay Youhnovski,Marcus Groettrup,and Michael Przybylski
第四部分 蛋白质组学定量技术
11. 基于液相色谱质谱联用的定量蛋白质组学 189
Michael W.Linscheid,Robert Ahrends,Stefan Pieper,and Andreas Kuhn
12. 胶内酶切同位素标记蛋白质技术用于相对定量 207
Carla Schmidt and Henning Urlaub
13.电喷雾质谱技术用于血浆蛋白质组定量分析 227
Hong Wang and Sam Hanash
第五部分 质谱数据解读
14. 算法和数据库 245
Lennart Martens and Rolf Apweiler
15. 鸟枪法蛋白质鉴定和质谱法蛋白质定量 261
Bingwen Lu,Tao Xu,Sung Kyu Park,and John R.Yates III
第六部分 蛋白质修饰分析
16. 脑蛋白氧化修饰蛋白质组学鉴定 291
Rukhsana Sultana,Marz ia Perluigi,and D.Allan Butter field
17.同位素标记、磷酸化多肽亲和富集技术用于液相色谱-串联质谱蛋白质组分析 303
Uma Kota,Ko-yi Chien,and Michael B.Goshe
第七部分 亚细胞蛋白质组学
18. 细胞器蛋白质组学:天然蛋白胶内酶切筛选减少样本的复杂性 325
Veronika Reisinger and Lutz A.Eichacker
19. 聚合物双水相系统逆流分配法分离神经系统质膜 335
Jens Schindler and Hans Gerd NothWang
20. 液相色谱串联质谱技术进行拟南芥蛋白质组分析的质膜蛋白富集和制备 341
Srijeet K.Mitra,Steven D.Clouse,and Michael B.Goshe
第八部分 蛋白质相互作用分析
21.Strep/FLAG(SF)-串联亲和纯化标签用于哺乳动物细胞蛋白质复合物串联亲和纯化 359
Christian Johannes Gloeckner,Karsten Boldt,Annette Schumacher,and Marius Ueffing
22. 顺序多肽亲和纯化系统分离和鉴定大肠杆菌蛋白质复合物 373
Mohan Babu,Gareth Butland,Oxana Pogoutse,Joyce Li,Jack F.Greenblatt,and AndreWEmili
23. 生物信息学方法分析蛋白质相互作用 401
Beate Kruger and Thomas Dandekar
索引 433
(宋凯 张春秀 译)
Contents
Preface. v
Contributors. ix
PART I INTRODUCTION
1. Introduction to Proteomics 3
Friedrich Lottspeich
PART II ELECTROPHORETIC SEPARATIONS
2. High-Resolution Two-Dimensional Electrophoresis 13
Walter Weiss and Angelika Gorg
3. Non-classical 2-D Electrophoresis 33
Jacqueline Burre,Ilka Wittig,and Hermann Sch?gger
4. Protein Detection and Quantitation Technologies for Gel-Based Proteome Analysis 59
Walter Weiss,Florian Weiland,and Angelika Gorg
PART III MASS SPECTROMETRY AND TANDEM MASS SPECTROMETRY APPLICATIONS
5. MALDI MS 85
Rainer Cramer
6. Capillary Electrophoresis Coupled to Mass Spectrometry for Proteomic Profiling of Human Urine and Biomarker Discovery 105
Petra Zurbig,Eric Schiffer,and Harald Mischak
7. A Newcomer’s Guide to Nano-Liquid-Chromatography of Peptides 123
Thomas Frohlich and Georg J. Arnold
8. Multidimensional Protein Identification Technology 143
Katharina Lohrig and Dirk Wolters
9. Characterization of Platelet Proteins Using Peptide Centric Proteomics 155
Oliver Simon,Stefanie Wortelkamp,and Albert Sickmann
10. Identification of the Molecular Composition of the 20S Proteasome of Mouse Intestine by High-Resolution Mass Spectrometric Proteome Analysis 173
Reinhold Weber,Regina Preywisch,Nikolay Youhnovski,Marcus Groettrup,and Michael Przybylski
PART IV QUANTITATIVE PROTEOMICS
11. Liquid Chromatography-Mass Spectrometry-Based Quantitative Proteomics. 189
Michael W. Linscheid,Robert Ahrends,Stefan Pieper,and Andreas Kuhn
12. iTRAQ-Labeling of In-Gel Digested Proteins for Relative Quantification 207
Carla Schmidt and Henning Urlaub
13. Electrospray Mass Spectrometry for Quantitative Plasma Proteome Analysis 227
Hong Wang and Sam Hanash
PART V INTERPRETATION OF MASS SPECTROMETRY DATA
14. Algorithms and Databases 245
Lennart Martens and Rolf Apweiler
15. Shotgun Protein Identification and Quantification by Mass Spectrometry 261
Bingwen Lu,Tao Xu,Sung Kyu Park,and John R. Yates III
PART VI ANALYSIS OF PROTEIN MODIFICATIONS
16. Proteomics Identification of Oxidatively Modified Proteins in Brain 291
Rukhsana Sultana,Marzia Perluigi,and D. Allan Butterfield
17. Isotope-Labeling and Affinity Enrichment of Phosphopeptides for Proteomic Analysis Using Liquid Chromatography-Tandem Mass Spectrometry 303
Uma Kota,Ko-yi Chien,and Michael B. Goshe
PART VII SUBCELLULAR PROTEOMICS
18. Organelle Proteomics:Reduction of Sample Complexity by Enzymatic In-Gel Selection of Native Proteins 325
Veronika Reisinger and Lutz A. Eichacker
19. Isolation of Plasma Membranes from the Nervous System by Countercurrent Distribution in Aqueous Polymer Two-Phase Systems 335
Jens Schindler and Hans Gerd Nothwang
20. Enrichment and Preparation of Plasma Membrane Proteins from Arabidopsis thaliana for Global Proteomic Analysis Using Liquid Chromatography-Tandem Mass Spectrometry 341
Srijeet K. Mitra,Steven D. Clouse,and Michael B. Goshe
PART VIII ANALYSIS OF PROTEIN INTERACTIONS
21. Tandem Affinity Purification of Protein Complexes from Mammalian Cells by the Strep/FLAG (SF)-TAP Tag 359
Christian Johannes Gloeckner,Karsten Boldt,Annette Schumacher,and Marius Ueffing
22. Sequential Peptide Affinity Purification System for the Systematic Isolation and Identification of Protein Complexes from Escherichia coli 373
Mohan Babu,Gareth Butland,Oxana Pogoutse,Joyce Li,Jack F. Greenblatt,and AndreWEmili
23. Bioinformatical Approaches to Detect and Analyze Protein Interactions. 401
Beate Kruger and Thomas Dandekar
Index 433