SELF ASSOCIATION OF PROTEINS (SIMSTEX): DEVELOPMENT & APPLICATON

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来源获得了主要资金， 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-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。 各种胰岛素及其类似物寡聚化的倾向可通过质谱方法研究，包括测量气相中寡聚物的相对丰度和H/D酰胺交换动力学。 氘吸收的动力学表现出良好的拟合时，交换酰胺被放置在三个动力学组：快，中间，和慢。在所研究的胰岛素中，r-人胰岛素具有较少的以中等速率交换的酰胺，而较多的以慢速率交换的酰胺，这与其较高的缔合程度一致雅阁。在溶液中。 我们采用PLIMSTEX（通过质谱法测定蛋白质配体相互作用，滴定，和H/D交换）以确定。溶液中的蛋白质/配体亲和力，到Det。自关联等。常数并将其应用于各种胰岛素类似物。 我们将这种适应称为SIMSTEX（自关联）。相互作用使用质谱，自滴定。和H/D交换）;它给出亲和力常数。这与文献结果相当。 SIMSTEX的结果表明，一些突变体（例如，GlnB 13）具有增加的自分泌倾向，可能减缓它们在体内的作用。 其他突变体（例如，赖脯氨酸和AspB 9）具有较低自缔合倾向，从而提供用于控制糖尿病的潜在的更快速作用的类似物。

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. The propensity of various insulins and their analogs to oligomerize can be investigated by mass spectrometric methods including measurement of the relative abundances of oligomers in the gas phase and the kinetics of H/D amide exchange. The kinetics of deuterium uptake show a good fit when the exchanging amides are placed in three kinetic groups: fast, intermediate, and slow. r-Human insulin, of the insulins investigated, has fewer amides that exchange at intermediate rates and more that exchange at slow rates, in accord with its higher extent of assocn. in soln. We adapted PLIMSTEX (protein ligand interactions by mass spectrometry, titrn., and H/D exchange) to det. protein/ligand affinities in soln., to det. self-assocn. equil. consts. for proteins, and to apply them to various insulin analogs. We term this adaptation SIMSTEX (self-assocn. interactions using mass spectrometry, self-titrn. and H/D exchange); it gives affinity consts. that compare well with the literature results. The results from SIMSTEX show that some mutants (e.g., GlnB13) have an increased tendency to self-assoc., possibly slowing down their action in vivo. Other mutants (e.g., lispro and AspB9) have lower propensities for self-assocn., thus providing potentially faster-acting analogs for use in controlling diabetes.