Analysis of protein-protein interactions in human plasma using non-denaturing two-dimensional electrophoresis and mass spectrometry

使用非变性二维电泳和质谱分析人血浆中的蛋白质-蛋白质相互作用

• 批准号: 16550077
• 负责人: MANABE Takashi
• 金额: $ 2.3万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16550077/
• 关键词: human plasma; plasma proteins; non-denaturing; two-dimensional gel electrophoresis; micro gel; mass spectrometry; MALDI-TOF MS; protein interaction; マイクロ; ESI-MS; MS; 生理的条件

Protein-protein interactions between human plasma proteins have been detected by micro two-dimensional electrophoresis (2-DE), comparing the patterns obtained under non-denaturing conditions and those obtained under partially denaturing conditions. The protein spots on the 2-DE gels, 157 spots on a non-denaturing micro 2-DE gel and 175 spots on a partially denaturing micro 2-DE gel, were subjected to structural analysis by mass spectrometry. For mass spectrometric analysis, matrix assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS) and database search (peptide mass fingerprinting, PMF) were employed. Integrating the information obtained, it was ascertained that structural information on human plasma proteins, including their status of complex-formation, can be analyzed. Although for the moment most of the protein researchers are using 2-DE under denaturing conditions, present study revealed that 2-DE under non-denaturing conditions will become more important by the advance of mass spectrometric methods in protein analysis.

用微双向电泳（2-DE）检测了人血浆蛋白之间的蛋白-蛋白相互作用，比较了在非变性条件下和部分变性条件下获得的模式。用质谱法对2-DE凝胶上的蛋白斑点、未变性微2-DE凝胶上的157个斑点和部分变性微2-DE凝胶上的175个斑点进行结构分析。质谱分析采用基质辅助激光飞行时间质谱法（MALDI-TOF MS）和数据库检索（多肽质量指纹图谱，PMF）。综合所获得的信息，确定可以分析人血浆蛋白的结构信息，包括它们的复合物形成状态。虽然目前大多数蛋白质研究人员使用的是变性条件下的2-DE，但目前的研究表明，随着质谱方法在蛋白质分析中的进步，非变性条件下的2-DE将变得更加重要。

项目成果

High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry

• DOI: 10.1002/elps.200410187
• 发表时间: 2005-03-01
• 期刊: ELECTROPHORESIS
• 影响因子: 2.9
• 作者: Jin, Y;Manabe, T
• 通讯作者: Manabe, T

Detection of activity and mass spectrometric identification of mouse liver carboxylesterase and aldehyde dehydrogenase separated by non-denaturing two-dimensional electrophoresis after extraction with detergents.

• DOI: 10.1016/j.bbapap.2005.02.011
• 发表时间: 2005-05
• 期刊: Biochimica et biophysica acta
• 作者: Y. Shimazaki;T. Manabe

Concentration and Separation of Low-Abundant Proteins in Human Plasma by Ammonium Sulfate Fractionation Followed by a Three-Step Electrophoresis Technique

通过硫酸铵分级和三步电泳技术浓缩和分离人血浆中的低丰度蛋白质

• 发表时间: 2004
• 期刊: J.Electrophoresis 48・2
• 作者: 向井 準;真鍋 敬
• 通讯作者: 真鍋 敬

The Most Advanced Analytical Methods (in Japanese)

最先进的分析方法（日语）

• 发表时间: 2004
• 作者: A.Kotani;S.Kojima;H.Hakamata;F.Kusu;Takashi Manabe
• 通讯作者: Takashi Manabe

Proteome Analysis of Cashmere

羊绒蛋白质组分析

• 发表时间: 2004
• 期刊: Anim.Sci.J. 75
• 作者: Michinari Yokohama;Takeshi Masuda;Takashi Amano;Hiroki Hirayama;Takashi Manabe
• 通讯作者: Takashi Manabe