Fremont D H, Anderson D H, Wilson I A, Dennis E A, Xuong N H
Department of Chemistry, University of California, San Diego, La Jolla 92093.
Proc Natl Acad Sci U S A. 1993 Jan 1;90(1):342-6. doi: 10.1073/pnas.90.1.342.
Phospholipase A2 (PLA2) from Indian cobra venom (Naja naja naja) was crystallized from ethanol in space group P4(3)2(1)2 in the presence of Ca2+. The x-ray crystal structure was determined to 2.3-A resolution by molecular replacement techniques using a theoretical model constructed from homologous segments of the bovine pancreatic, porcine pancreatic, and rattlesnake venom crystal structures. The structure was refined to an R value of 0.174 for 17,542 reflections between 6.0- and 2.3-A resolution (F > 2 sigma), including 148 water molecules. The 119-amino acid enzyme has an overall architecture strikingly similar to the other known PLA2 structures with regions implicated in catalysis showing the greatest structural conservation. Unexpectedly, three monomers were found to occupy the asymmetric unit and are oriented with their catalytic sites facing the pseudo-threefold axis with approximately 15% of the solvent accessible surface of each monomer buried in trimer contacts. The majority of the interactions at the subunit interfaces are made by residues unique to PLA2 sequences from cobra and krait venoms. The possible relevance of this unique trimeric structure is considered.
印度眼镜蛇(Naja naja naja)毒液中的磷脂酶A2(PLA2)在Ca2+存在的情况下,于空间群P4(3)2(1)2中从乙醇中结晶出来。通过分子置换技术,利用由牛胰、猪胰和响尾蛇毒液晶体结构的同源片段构建的理论模型,将X射线晶体结构解析到了2.3埃的分辨率。该结构在6.0至2.3埃分辨率(F > 2σ)下对17542个反射进行精修,R值为0.174,包括148个水分子。这个119个氨基酸的酶的整体结构与其他已知的PLA2结构惊人地相似,参与催化的区域显示出最大的结构保守性。出乎意料的是,发现三个单体占据不对称单元,并且它们的催化位点朝向伪三重轴,每个单体约15%的溶剂可及表面埋在三聚体接触中。亚基界面处的大多数相互作用是由眼镜蛇和金环蛇毒液PLA2序列特有的残基形成的。文中考虑了这种独特三聚体结构的可能相关性。