MECHANISTIC STRUCTURE-FUNCTION-INHIBITOR STUDIES

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 项目一：糖尿病和肥胖症的发病率正在以惊人的速度增长，影响着世界各地的数百万人。人胰腺α-淀粉酶(HPA；496A.A.‘S)是开发这两种疾病潜在疗法的独特且引人注目的靶点，因为它在淀粉消化中发挥核心作用。然而，此类疗法的成功开发依赖于对HPA的催化残基如何发挥作用的结构理解以及延长的结合裂隙中底物结合的方式。尽管我们已经成功地应用了定点突变技术并生长了野生型和变异型蛋白质的晶体，但由于使用我们自己的实验室X射线源(Rigaku RU-300)缺乏分辨率(~2.0？)，解释由底物和抑制剂形成的复合体的结构结果的成功有限。使用SSRL数据收集设施将极大地提高我们结构研究的分辨率，从而允许对这种机械复合体进行全面解释，并促进基于这些机械数据的新疗法的开发。项目II：令人惊讶的是，只有革兰氏阴性细菌含有六聚体II型柠檬酸合成酶，这种合成酶具有代谢调节的特殊性质。相比之下，其他生物体(包括人类)的二聚体柠檬酸合成酶是不受监管的。由于许多革兰氏阴性菌是危险的人类病原体，II型柠檬酸合成酶的特殊性质可能成为开发新型抗微生物药物的基础。开发这类抗微生物药物的关键是六聚体II型柠檬酸合成酶(CS)在其各种变构和代谢控制状态下的结构特征。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Project I: The incidence of diabetes and obesity is increasing at an alarming rate, affecting millions around the world. Human pancreatic alpha-amylase (HPA; 496 a.a.'s) is a unique and compelling target for the development of potential therapeutics for both these diseases, given its central role in starch digestion. However, successful development of such therapeutics is dependent on a structural understanding of how the catalytic residues of HPA function and the mode of substrate binding in the elongated binding cleft present. Although we have successfully applied site directed mutagenesis techniques and grown crystals of both wild-type and variant proteins, interpretation of the structural results of complexes formed by substrates and inhibitors has had limited success due to a lack of resolution (~2.0 ¿) using our home laboratory x-ray source (Rigaku RU-300). Access to the SSRL data collection facility will greatly enhance the resolutions of our structural studies, thereby allowing for a comprehensive interpretation of such mechanistic complexes and facilitating the development of novel therapeutics based on this mechanistic data. Project II: Surprisingly, only Gram-negative bacteria contain hexameric Type II citrate synthases that have the special property of being metabolically regulated. In contrast, the dimeric citrate synthases of other organisms (including humans) is unregulated. Since many Gram-negative bacteria are dangerous human pathogens, the special properties of Type II citrate synthases could form a basis for the development of novel anti-microbials. Key to the development of such anti-microbials is the structural characterization of a hexameric Type II citrate synthase (CS) in its various allosteric and metabolically controlled states.