X-RAY CRYSTALLOGRAPHIC STUDIES OF METAL TRAFFICKING PROTEINS AND METALLOENZYMES

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。从我们之前的提议中获得的束流时间导致了金属运输蛋白和金属酶的结构的确定。金属转运蛋白的结构包括铜伴侣蛋白Hah1和CCS，细菌抗铜蛋白PcoC，以及酵母核苷酸还原酶Y2和Y4蛋白之间的异源二聚体复合体。酶结构包括β-内酰胺合成酶和核糖核苷酸还原酶R2的突变体。我们建议继续我们对CCS、PcoC、Y2、Y4和R2的研究。此外，我们还在进行与β-内酰胺合成酶相关的CARA的结构测定。最后，我们得到了颗粒甲烷单加氧酶(PMMO)的衍射型晶体，这是一种膜结合的酶，它利用催化铜中心将甲烷氧化为甲醇，并计划通过铜多波长反常色散(MAD)位相来解决这种结构。

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. Beam time obtained from our previous proposal has led to the determination of structures of metal trafficking proteins and metalloenzymes. Structures of metal trafficking proteins include the copper chaperones Hah1 and CCS, the bacterial copper resistance protein PcoC, and the heterodimeric complex between the yeast ribonucleotide reductase Y2 and Y4 proteins. Enzyme structures include beta-lactam synthetase and mutants of ribonucleotide reductase R2. We propose to continue our studies of CCS, PcoC, Y2, Y4, and R2. In addition, we are pursuing the structure determination of CarA, which is related to beta-lactam synthetase. Finally, we have obtained diffracting crystals of particulate methane monooxygenase (pMMO), a membrane-bound enzyme that uses a catalytic copper center to oxidize methane to methanol, and plan to solve this structure by copper multiwavelength anomalous dispersion (MAD) phasing.