STRUCTURAL ANALYSIS OF SPINACH ACYL CARRIER PROTEIN

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。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。所列机构为中心，不一定是研究者所在机构。本项目的目的是确定菠菜酰基载体蛋白（ACP）的脱辅基、全酰基和酰基三种形式的三维溶液结构。提出了E. coliACP已报道，然而，结构是在没有使用15 N和13 C标记蛋白的情况下完成的。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相互转化。已经开发了产生大量（350 mg）15 N和13 C标记的菠菜apo-ACP的方法，所述apo-ACP然后可以在体外转化为全和酰基形式。通过NMR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种构象之间的相互转换问题。

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.