METALLOPROTEINS FOR NITRIC OXIDE TRANSPORT AND OTHER ACTIVITIES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这是一个项目提案，用于研究五种名为亚硝酚的一氧化氮(NO)转运蛋白，以及一种名为CueO的铜伴侣蛋白。NO是高等生物体中普遍存在的一种信号分子，对医学和基础科学具有重要意义。亚硝酚来自两种吸血昆虫，长尾罗非鱼(Rhodnius prolixus，rNP1，rNP2，rNP4)和Cimex lectularius(CNP)，为了解蛋白质中的非血红素相互作用提供了最好的机会。我们有rNP1、rNP2和rNP4的晶体结构，以及良好的CNP和CueO晶体。建议用于rNP4(至0.8°)的超高分辨率研究、与所有硝基酚及其突变体的NO络合物的结构测定、与在晶体中经历较大构象变化的NP4的冷冻捕获实验以及对CNP和CueO的MAD实验。由于铜K-α辐射产生的光解或光还原效应，测定NO复杂结构需要在同步加速器上使用较短的波长。

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.