来自维氏硫杆菌的喹蛋白甲胺脱氢酶的纯化、结晶及初步X射线研究。

Purification, crystallization and preliminary X-ray investigation of quinoprotein methylamine dehydrogenase from Thiobacillus versutus.

作者信息

Vellieux F M, Frank J, Swarte M B, Groendijk H, Duine J A, Drenth J, Hol W G

出版信息

Eur J Biochem. 1986 Jan 15;154(2):383-6. doi: 10.1111/j.1432-1033.1986.tb09409.x.

DOI:10.1111/j.1432-1033.1986.tb09409.x

Abstract

The enzyme methylamine dehydrogenase or primary-amine:(acceptor) oxidoreductase (deaminating) (EC 1.4.99.3) was purified from the bacterium Thiobacillus versutus to homogeneity, as judged by polyacrylamide gel electrophoresis. The native enzyme has a Mr of 123 500 and contains four subunits arranged in a alpha 2 beta 2 configuration, the light and heavy subunits having a Mr of 12900 and 47500 respectively. The isoelectric point is 3.9. The purified enzyme was crystallized from 37--42% saturated ammonium sulphate in 0.1 M sodium acetate buffer, pH 5.0. The space group is P3(1)21 or P3(2)21, with one alpha 2 beta 2 molecule in the asymmetric unit. The cell dimensions are: a = b = 13.01 nm; c = 10.40 nm. The X-ray diffraction pattern extends to at least 0.25-nm resolution.

摘要

通过聚丙烯酰胺凝胶电泳判断，从维氏硫杆菌中纯化得到了甲胺脱氢酶或伯胺：（受体）氧化还原酶（脱氨基）（EC 1.4.99.3），达到了同质纯。天然酶的相对分子质量为123500，包含以α₂β₂构型排列的四个亚基，轻亚基和重亚基的相对分子质量分别为12900和47500。其等电点为3.9。纯化后的酶在0.1M醋酸钠缓冲液（pH 5.0）中，由37%-42%饱和度的硫酸铵结晶得到。空间群为P3(1)21或P3(2)21，不对称单元中有一个α₂β₂分子。晶胞参数为：a = b = 13.01 nm；c = 10.40 nm。X射线衍射图谱的分辨率至少达到0.25 nm。