Balasubramanian Moovarkumudalvan, Moorthy Ponnuraj Sathya, Neelagandan Kamariah, Ponnuswamy Mondikalipudur Nanjappa Gounder
Centre of Advanced Study in Crystallography and Biophysics, University of Madras, Guindy Campus, Chennai - 600 025, India.
Protein Pept Lett. 2009;16(2):213-5. doi: 10.2174/092986609787316216.
Hemoglobin is a tetrameric, iron-containing metalloprotein, which plays a vital role in the transportation of oxygen from lungs to tissues and carbon dioxide back to lungs. Though good amount of work has already been done on hemoglobins, the scarcity of data on three dimensional structures pertaining to low oxygen affinity hemoglobins from mammalian species, motivated our group to work on this problem specifically. Herein, we report the preliminary crystallographic analysis of buffalo hemoglobin, which belongs to low oxygen affinity species. The buffalo blood was collected, purified by anion exchange chromatography and crystallized with PEG 3350 using 50mM phosphate buffer at pH 6.7 as a precipitant by hanging drop vapor diffusion method. Data collection was carried out using mar345dtb image plate detector system. Buffalo hemoglobin crystallizes in orthorhombic space group P2(1)2(1)2(1) with one whole biological molecule (alpha2beta2) in the asymmetric unit with cell dimensions a=63.064A, b=74.677A, c=110.224A.
血红蛋白是一种含四个亚基的含铁金属蛋白，在将氧气从肺部运输到组织以及将二氧化碳运回肺部的过程中起着至关重要的作用。尽管已经对血红蛋白进行了大量研究，但关于哺乳动物低氧亲和力血红蛋白三维结构的数据却很匮乏，这促使我们团队专门研究这个问题。在此，我们报告水牛血红蛋白的初步晶体学分析，水牛血红蛋白属于低氧亲和力物种。采集水牛血液，通过阴离子交换色谱法进行纯化，并采用悬滴气相扩散法，以pH 6.7的50mM磷酸盐缓冲液作为沉淀剂，用聚乙二醇3350使其结晶。使用mar345dtb图像板探测器系统进行数据收集。水牛血红蛋白在正交晶系空间群P2(1)2(1)2(1)中结晶，不对称单元中有一个完整的生物分子（α2β2），晶胞参数为a = 63.064Å，b = 74.677Å，c = 110.224Å。
