Crystallization and preliminary X‐ray analysis of outer membrane phospholipase A from Escherichia coli

FEBS Letters

Blaauw, Mieke²  Kalk, Kor H.²  Dekker, Niek¹  Verheij, Hubertus M.¹  Dijkstra, Bauke W.² 

[1] Centre for Biomembranes and Lipid Enzymology, Department of Enzymology and Protein Engineering, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands;Laboratory of Biophysical Chemistry and BIOSON Research Institute, Department of Chemistry, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands

关键词: Outer membrane phospholipase;    Membrane protein crystallization;    X-Ray crystallography;    E. coli;    Bis-Tris;    bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane;    MPD;    2-methyl-2;    4-pentanediol;    OMPLA;    outer membrane phospholipase A (EC 3.1.1.32);    PEG 400;    polyethyleneglycol 400;    V M;    Matthews coefficient;   

DOI  :  10.1016/0014-5793(95)01002-V

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The outer membrane phospholipase A (OMPLA) of Escherichia coli is one of the few integral outer membrane proteins displaying enzymatic activity. It is encoded as a mature protein of 269 amino acids preceded by a signal sequence of 20 amino acids. There is no sequence homology with water-soluble lipases and phospholipases. Crystals of the mature enzyme were obtained at 22°C from 24–28% (vlv) 2-methyl-2,4-pentanediol in Bis-Tris buffer, pH 5.9–6.0, with 1 mM calcium chloride and 1.5% (w/v) β-octylglucoside. They have the symmetry of the trigonal spacegroup P3₁21 (or P3₂21) with cell dimensions of ). Native crystals diffract to a resolution of 2.6 Å.

【 授权许可】

Unknown