IMPROVED METHODS FOR THE ANALYSIS OF PROTEIN COMPLEXES

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们并列了几种方法，以提高复杂蛋白质混合物（5-50个蛋白质）中蛋白质的鉴定。因此，我们比较了直接消化蛋白质混合物，然后用色氨酸肽液相色谱-质谱法鉴定色谱组分中的肽，以及凝胶内分离蛋白质，然后用完整的凝胶切片和质谱法鉴定凝胶片中的每一片色氨酸肽。我们的总凝胶切片技术的优化导致了解决合作研究部分中描述的具有挑战性的生物学问题的改进方法。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have juxtaposed several methods in an effort to improve the identification of proteins in complex protein mixtures (5-50 proteins.). Thus, we have compared direct digestion of protein mixtures followed by tryptic peptide liquid chromatography-mass spectrometric identification of peptides in chromatographic fractions with in-gel separation of proteins followed by complete gel slicing and mass spectrometric identification of tryptic peptides in every piece of a gel slice. Optimization of our total gel slicing technique is leading to an improved method for solving challenging biological problems described in the Collaborative Research section.