Mechanism of Nickel Incorporation into Urease

对酶金属中心的生物合成了解甚少。本研究以脲酶为模型，旨在了解金属与蛋白质基质结合的一般机制。脲酶作为载脂蛋白酶合成，需要一个辅助因子来形成其新颖的双镍活性位点。辅助因素的性质是未知的；然而，两个新发现的基因似乎在这一过程中发挥了作用。这些基因的突变将被产生，突变对载脂蛋白的影响将被确定。辅助蛋白将被分离出来并进行细胞定位、Ni结合和与载脂蛋白脲酶相互作用的表征。这项研究将增加我们对金属蛋白如何形成的理解，以及我们对生命系统中镍的生物化学的理解。

The biosynthesis of enzyme metallocenters is very poorly understand. This proposal is aimed at understanding the general mechanism for incorporation of the metal into the protein matrix using urease as a model. Urease, synthesized as an apo-enzyme, requires an accessory factor for the formation of its novel, bi- nickel active site. The nature of the accessory factor is unknown; however, two newly identified genes appear to play a role in this process. Mutants of these genes will be generated and the effects of the mutations on the apo-protein will be determined. The accessory protein (s) will be isolated and characterized for cellular location, Ni binding and interaction with apo-urease. The research will increase our understanding of how metalloproteins are formed, and our understanding of the biochemistry of nickel in living systems.