ISOTOPE SIGNATURE BASED IDENTIFICATION OF CROSSLINKED PEPTIDES BY MS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蛋白质结构和蛋白质-蛋白质相互作用的知识对于我们理解生物过程至关重要。获得这种知识往往是一个严峻的挑战。蛋白质交联和质谱（MS）的最新进展显示出对这一领域做出贡献的巨大潜力。在这里，我们正在开发一种新的方法，以快速，准确地识别交联肽的基础上，其独特的同位素签名时，在H218 O的存在下消化。该方法克服了对特殊合成的交联剂和/或其他技术所需的多次MS运行的需要。我们将通过分析已知结构的蛋白质/复合物来验证我们的方法。计算技术Rosetta将被修改，以验证对公布的原子结构观察到的交联。

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. Knowledge of protein structures and protein-protein interactions is essential for our understanding of biological processes. Obtaining such knowledge often presents a severe challenge. Recent advances in protein crosslinking and mass spectrometry (MS) have shown significant potential to contribute to this area. Here we are developing a novel method to rapidly and accurately identify crosslinked peptides based on their unique isotope signature when digested in the presence of H218O. This method overcomes the need for specially synthesized crosslinkers and/or multiple MS runs required by other techniques. We will validate our method by analyzing proteins/complexes of known structure. The computational technique Rosetta will be modified to validate crosslinks observed against published atomic structures.