THIOREDUCTASE EXPRESSED IN SF21 CELLS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目涉及一种不寻常的蛋白质二硫键异构酶GILT，它在溶酶体的酸性条件下工作。它参与mhc II类成熟，也在病原体劫持细胞中发挥关键作用。我们有硒代甲硫氨酸取代的两个双胍突变体的晶体，我们想用它们来定相。这些晶体可以在我们的家庭源上表现得比3A更好，我们期望在同步加速器源上有更好的分辨率。我们有几个预先筛选的晶体和~20，我们也想筛选/收集上GILT的结构研究将有助于我们阐明蛋白质的酶机制，以及它如何在溶酶体的酸性pH值下工作。此外，这项工作将奠定基础，了解其参与致病过程的分子水平。

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. The project involves an unusual protein disulfide isomerase, GILT, that works at acidic conditions in the lysosome. It is involved in mhc class II maturation and also plays a key role in the hi-jacking of cells by pathogens. We have selenomethionine substituted crystals of two differenet mutants with which we would like to use for phasing. These crystals can diffract to better than 3A on our home source and we expect better resolution at a synchrotron source. We have several pre-screened crystals and ~20 that we would also like to screen/collect on.The structural studies on GILT will help us to elucidate the enzymatic mechanism of the protein, and how it works at the acidic pH of the lysosome. Furthermore, this work will lay the basis to understand its involvement in pathogenic processes on a molecular level.