ACTIVE SITE ARCHITECTURE OF DISPERSIN B, A BETA-HEXOSAMINIDASE

分散素 B（一种 β-氨基己糖苷酶）的活性位点结构

• 批准号: 7721326
• 负责人: NARAYANAN RAMASUBBU
• 金额: $ 1.71万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721326
• 关键词: Acetates; Acetylglucosamine; Active Sites; Architecture; Binding; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Data; Enzymes; Funding; Grant; Hydrolysis; Institution; Research; Research Personnel; Resolution; Resources; Role; Source; Structure; United States National Institutes of Health; base; beta-n-acetylhexosaminidase; mutant; prevent

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. We have solved the crystal structure of dispersin B at 2.0 A resolution. Our current project focuses on the detremination of active ste residues and their role in hydrolysis. We have made several mutants of this enzyme based on the sequence similarity of the enzyme with other beta-hexosaminidases. We would now like to identify the various subites present in this molecule. Our initial efforts will focus on the co-crystallization of N-acetylglucosamine anf the mutant enzymes. Our initial stuidies have produced crystals that diffract upto 2.7A. However, the active site has a bound acetate that prevents N-acetylglucosamine to occupy the active only partially. Since the crystals are small, we feel that collecting data at the beam line should increase the resolution. The high resolution data would be of great use for our studies.

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。 我们已经解决了分散素B在2.0 A分辨率的晶体结构。我们目前的项目集中在活性ste残留物的测定及其在水解中的作用。基于该酶与其他β-氨基己糖苷酶的序列相似性，我们已经制备了该酶的几种突变体。我们现在想识别该分子中存在的各种亚基。我们最初的努力将集中在N-乙酰葡糖胺和突变酶的共结晶。我们最初的研究已经产生了晶体，其折射率高达2.7A。然而，活性位点具有结合的乙酸盐，其阻止N-乙酰葡糖胺仅部分地占据活性。由于晶体很小，我们觉得在光束线上收集数据应该会提高分辨率。这些高分辨率的数据将对我们的研究有很大的帮助。