SELF ASSOCIATION OF PROTEINS (SIMSTEX): DEVELOPMENT & APPLICATON

蛋白质自缔合 (SIMSTEX)：开发

• 批准号: 7355219
• 负责人: MICHAEL L GROSS
• 金额: $ 1.07万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7355219
• 关键词: SELF; ASSOCIATION; PROTEINS; SIMSTEX; DEVELOPMENT

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. Protein-protein interactions are crucial for many biological functions in living systems Disruptions in these interactions can lead to various diseases including metabolic disorders, cancer and infections. We are interested in developing mass spectrometric methods that take advantage of the high sensitivity and specificity of mass spectrometry, combined with H/D exchange and mathematical modeling to study self association. Insulin, a 51-amino acid protein is used in the treatment of diabetes. The tendency of human insulin to self associate to form hexamers in solution when reduced by mutating key residues gives faster-acting insulin analogs. Self association Interactions using Mass Spectrometry, self Titration and amide EXchange (SIMSTEX), a technique we developed to obtain affinity constants for oligomerization from titration data was applied to self association of insulin and insulin analogs. The affinity constants, we calculated, agree within a factor of 20 to the literature values for human and lispro insulins. The trends we observed for insulin mutants, also, agree well with literature reports, establishing SIMSTEX as a reliable technique to study self association.

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来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定为研究者机构。蛋白质-蛋白质相互作用对于生命系统中的许多生物功能至关重要。这些相互作用的破坏可导致各种疾病，包括代谢紊乱、癌症和感染。我们有兴趣开发质谱方法，利用质谱的高灵敏度和特异性，结合H/D交换和数学建模来研究自缔合。 胰岛素是一种由51个氨基酸组成的蛋白质，用于治疗糖尿病。当通过突变关键残基而减少时，人胰岛素在溶液中自缔合形成六聚体的趋势产生更快作用的胰岛素类似物。使用质谱、自滴定和酰胺交换的自缔合相互作用（SIMSTEX），我们开发的一种从滴定数据获得寡聚化亲和常数的技术被应用于胰岛素和胰岛素类似物的自缔合。我们计算的亲和常数与人胰岛素和赖脯胰岛素的文献值在20倍以内一致。我们观察到的胰岛素突变体的趋势，也同意与文献报道，建立SIMSTEX作为一个可靠的技术来研究自我协会。