Mirkin Nurit, Jaconcic Jean, Stojanoff Vivian, Moreno Abel
Hunter College, City University of New York, New York, New York, USA.
Proteins. 2008 Jan 1;70(1):83-92. doi: 10.1002/prot.21452.
Cytochrome c is one of the most studied proteins probably due to its electron-transfer properties in aerobic and anaerobic respiration. Particularly, cytochrome c from bovine heart is a small protein, M(r) 12,230 Da, globular (hydrodynamic diameter of 3.4 nm), soluble in different buffer solutions, and commercially available. Despite being a quite well-studied protein and relatively easy to manipulate from the biochemical and electrochemical viewpoint, its 3D structure has never been published. In this work, the purification, crystallization and 3D structure of one of the cytochrome c isoforms is presented to 1.5 A resolution. It is also shown how the presence of isoforms made both the purification and crystallization steps difficult. Finally, a new approach for protein electrocrystallization and design of biosensors is presented.
细胞色素c可能是研究最为深入的蛋白质之一，这大概归因于其在有氧和无氧呼吸中的电子传递特性。特别是来自牛心的细胞色素c，它是一种小蛋白，分子量为12,230道尔顿，呈球状（流体动力学直径为3.4纳米），可溶于不同的缓冲溶液，且有商业产品供应。尽管它是一种研究得相当透彻的蛋白质，并且从生化和电化学角度相对易于操作，但其三维结构从未发表过。在这项工作中，展示了一种细胞色素c同工型的纯化、结晶及其三维结构，分辨率达到1.5埃。还表明了同工型的存在如何使纯化和结晶步骤变得困难。最后，提出了一种蛋白质电结晶和生物传感器设计的新方法。
