嗜热脂肪芽孢杆菌3-磷酸甘油酸激酶的纯化、结晶及初步X射线分析

Purification, crystallization and preliminary X-ray analysis of the 3-phosphoglycerate kinase from Bacillus stearothermophilus.

作者信息

Davies G J, Gamblin S J, Littlechild J A, Watson H C

机构信息

Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.

出版信息

J Mol Biol. 1992 Oct 20;227(4):1263-4. doi: 10.1016/0022-2836(92)90538-u.

DOI:10.1016/0022-2836(92)90538-u

PMID:1433299

Abstract

As part of a programme investigating the molecular basis of thermal stability in proteins we have isolated and characterized the thermally stable 3-phosphoglycerate kinase (PGK) from Bacillus stearothermophilus NCA 1503. The B. stearothermophilus PGK has been crystallized in a form suitable for X-ray diffraction analysis. Crystals which diffract to greater than 1.8 A resolution have been grown in the presence of the nucleotide substrate, MgATP, using polyethylene glycol (PEG 600) as a precipitant. The best crystals have been obtained using "seeding" techniques and are monoclinic, space group P2(1), with cell dimensions a = 40.5 A, b = 74.0 A, c = 68.5 A and beta = 99.8 degrees.

摘要

作为一项研究蛋白质热稳定性分子基础的计划的一部分，我们从嗜热脂肪芽孢杆菌NCA 1503中分离并鉴定了热稳定的3-磷酸甘油酸激酶（PGK）。嗜热脂肪芽孢杆菌PGK已结晶成适合X射线衍射分析的形式。在核苷酸底物MgATP存在下，使用聚乙二醇（PEG 600）作为沉淀剂，已生长出衍射分辨率大于1.8 Å的晶体。使用“接种”技术获得了最佳晶体，其为单斜晶系，空间群P2(1)，晶胞参数a = 40.5 Å，b = 74.0 Å，c = 68.5 Å，β = 99.8°。