STRUCTURAL ANALYSIS OF SPINACH ACYL CARRIER PROTEIN

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。本项目的目标是确定菠菜酰基载体蛋白(ACP)以apo、holo和acyl形式存在的三维溶液结构。已经报道了一种大肠杆菌ACP的溶液结构，然而，该结构是在没有使用15N和13C标记的蛋白质的情况下完成的。此外，据报道，该蛋白质在1秒的时间尺度上在两种结构形式之间进行相互转换。已经开发了大量(350 Mg)15N和13C标记的菠菜APO-ACP的生产方法，然后这些APO-ACP可以在体外转化为全息和酰基形式。通过用核磁共振表征这三种形式，我们打算解决每种形式之间发生的结构变化以及多个构象之间的相互转化问题。

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. The goal of this project is the determine the three dimensional solution structure of spinach acyl carrier protein (ACP) in the apo, holo and acyl forms. A solution structure for E. coli ACP has been reported, however, the structure was completed without using 15N and 13C labeled protein. Additionally, it was reported that the protein interconverts between two structural forms on a 1 second time-scale. Methods have been developed to produce large quantities (350 mg) of 15N and 13C labelled spinach apo-ACP which can then be converted to holo and acyl forms in vitro. By characterizing these three forms by NMR we intend to address the structural changes that occur between each form and also the issue of interconversion between multiple conformations.