METALLOPROTEINS FOR NITRIC OXIDE TRANSPORT AND OTHER ACTIVITIES

该子项目是利用 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来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 这是一个研究五种一氧化氮（NO）转运蛋白（称为nitrophorins）和一种铜伴侣蛋白（称为CueO）的计划提案。 NO是高等生物体中普遍存在的信号分子，是医学和基础科学的重要研究对象。 硝化蛋白来自两种吸血昆虫，Rhodnius prolixus（rNP 1，rNP 2，rNP 4）和Cimex lectularius（cNP），并为了解蛋白质中的NO-血红素相互作用提供了最好的机会。 我们具有rNP 1、rNP 2和rNP 4的晶体结构，以及cNP和CueO的良好晶体。 该提案是对rNP 4（至0.8 μ m）的100倍分辨率研究，NO与所有nitrophorins及其突变形式的复合物的结构测定，NP 4的冷冻捕获实验，其在晶体中经历了大的构象变化，以及cNP和CueO的MAD实验。 在同步加速器可用的较短的波长所需的NO络合物结构的测定，由于光解离或光还原的影响，发生与铜K-α辐射。

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. This is a program proposal for the study of five nitric oxide (NO) transport proteins called nitrophorins, and for a copper chaperone protein called CueO. NO is a ubiquitous signaling molecule in higher organisms and is of interest to medical and fundamental sciences. The nitrophorins are from two blood-sucking insects, Rhodnius prolixus (rNP1, rNP2, rNP4) and Cimex lectularius (cNP), and provide the best opportunity to date for understanding NO-heme interactions in proteins. We have crystal structures of rNP1, rNP2, and rNP4, and good crystals of cNP and CueO. The proposal is for ultrahigh resolution studies of rNP4 (to 0.8 ¿), structure determinations of NO complexes with all of the nitrophorins and their mutant forms, freeze trapping experiments with NP4, which undergoes a large conformational change in the crystal, and MAD experiments for cNP and CueO. The shorter wavelengths available at the synchrotron are required for the NO complex structure determinations, due to photodissociation or photoreduction affects that occur with Cu K-alpha radiation.