IMPROVED METHODS FOR THE ANALYSIS OF PROTEIN COMPLEXES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们已经并置了几种方法，以提高复杂蛋白质混合物（5-50种蛋白质）中蛋白质的鉴定。因此，我们已经比较了直接消化蛋白质混合物，然后胰蛋白酶肽液相色谱-质谱鉴定的肽在色谱馏分与凝胶内分离的蛋白质，然后由完整的凝胶切片和质谱鉴定的胰蛋白酶肽在每一块的凝胶切片。我们的总凝胶切片技术的优化导致了一种改进的方法，用于解决合作研究部分中描述的具有挑战性的生物学问题。

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.