日本蝮蛇毒液酸性磷脂酶A2的氨基酸序列修正、晶体化及初步X射线衍射分析

Revised amino acid sequence, crystallization, and preliminary x-ray diffraction analysis of acidic phospholipase A2 from the venom of Agkistrodon halys blomhoffii.

作者信息

Tomoo K, Ohishi H, Ishida T, Inoue M, Ikeda K, Aoki Y, Samejima Y

机构信息

Department of Physical Chemistry, Osaka University of Pharmaceutical Sciences, Japan.

出版信息

J Biol Chem. 1989 Feb 25;264(6):3636-8.

PMID:2914966

Abstract

The complete amino acid sequence of acidic Agkistrodon halys blomhoffii phospholipase A2 has been redetermined by a combination of manual and automatic Edman degradations. Acidic A. halys blomhoffi phospholipase is a single polypeptide chain consisting of 122 amino acids and is highly homologous in sequence with corresponding regions of phospholipase A2 from a variety of sources. Prism crystals of acidic A. halys blomhoffii phospholipase have been reproducibly grown from 2-methyl-2,4-pentanediol solution adjusted to pH 8.0 with 50 mM Tris-HCl buffer in the presence of 10 mM CaCl2. The crystals belong to space group P6(1)22 or P6(5)22 with hexagonal unit cell dimensions of a = b = 76.22 A and C = 76.56 A. One molecule occupies the asymmetric unit of the crystal. The diffraction extends to at least 2.5 A.

摘要

通过手动和自动埃德曼降解相结合的方法，重新测定了酸性蝮蛇（Agkistrodon halys blomhoffii）磷脂酶A2的完整氨基酸序列。酸性蝮蛇磷脂酶是由122个氨基酸组成的单条多肽链，其序列与多种来源的磷脂酶A2的相应区域高度同源。酸性蝮蛇磷脂酶的棱柱晶体已在含有10 mM氯化钙的情况下，从用50 mM Tris-HCl缓冲液调节至pH 8.0的2-甲基-2,4-戊二醇溶液中可重复生长出来。这些晶体属于空间群P6(1)22或P6(5)22，六方晶胞参数为a = b = 76.22 Å，c = 76.56 Å。一个分子占据晶体的不对称单元。衍射至少延伸到2.5 Å。