RABBIT MUSCLE ALDOLASE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 醛缩酶是从原核生物到哺乳动物的所有生物体中普遍存在的酶。它们有能力裂解碳-碳键。它们在糖酵解中的作用最为人所知，在糖酵解中，1，6-二磷酸果糖(FBP)醛缩酶将FBP分解为二羟丙酮-磷酸(DHAP)和甘油脱氢-3-磷酸。由于缺乏真正的反应中间体晶体结构，对残基的催化作用一直受到阻碍。 本课题的目的是研究兔肌醛缩酶的催化机理，该酶是一类1，6-二磷酸果糖缩醛酶。我们计划用天然和突变的醛缩酶晶体进行底物浸泡实验，以深入了解与催化有关的活性部位的残基。通过组合不同的浸泡时间，我们想要捕捉反应过程中的催化反应中间体，这意味着3个共价中间体的形成。 在过去的几年里，时间分辨结晶学已经成功地应用于用配体或底物浸泡的兔肌醛缩酶晶体。利用这项技术仍有可能进一步深入了解醛缩酶的催化机理。

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. Aldolase are ubiquitous enzymes found in all organisms, from prokaryotes to mammals. They have the ability to cleave carbon-carbon bonds. Their role is best known in glycolysis where fructose-1,6-bisphosphate (FBP) aldolase cleaves FBP to dihydroxyacetone-phosphate (DHAP) and glyceraldehy-3-phosphate. The catalytic role assigment to residues has beeen hampered by the lack of genuine reaction intermediate crystallographic structures. The aim of this project is to study the catalytic mechanism of the rabbit muscle aldolase, which is a class I fructose-1,6-bisphosphate aldolase. We are planning to use subtrate soaking experiments with native and mutant aldolase crystals to get insight about residues of the active site implicated in the catalysis. By using a combination of diverse soaking time, we want to trap catalytic reaction intermdiates among the reaction scheme, which is known to implicate the formation of 3 covalent intermediates. Time-resolved crystallography applied to rabbit muscle aldolase crystal soaked with ligands or substrates have been successfully used in the past years. Further insight into the catalytic mechanism of aldolase still possible by using this technique.