CRYSTALLIZATION AND CHARACTERIZATION OF COLICIN E1 CHANNEL-FORMING POLYPEPTIDES

被引：16

作者：

ELKINS, PA ^{[1
]}

SONG, HY ^{[1
]}

CRAMER, WA ^{[1
]}

STAUFFACHER, CV ^{[1
]}

机构：

[1] PURDUE UNIV, DEPT BIOL SCI, W LAFAYETTE, IN 47907 USA

来源：

PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS | 1994年 / 19卷 / 02期

关键词：

X-RAY CRYSTALLOGRAPHY; MEMBRANE PROTEIN; ION CHANNELS;

D O I：

10.1002/prot.340190208

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Crystals of the channel-forming domain of colicin E1 from E. coli were grown by vapor diffusion at pH 6.4 and higher pH values. Cleavage of the colicin molecule with trypsin or thermolysin produced two of the pore-forming polypeptides used in these experiments. The third polypeptide was purified from a constructed plasmid that overexpresses only the C-terminal domain of colicin E1. Polypeptide crystals are tetragonal with space group I4, have one monomer in the asymmetric unit, and diffract to 2.2-2.4 Angstrom. Unit cell parameters for the tryptic and thermolytic polypeptides are a = 102.9 Angstrom and c = 35.6 Angstrom. Crystals of the overexpressed polypeptide have unit cell parameters of a = 87.2 Angstrom and c = 59.1 Angstrom. The crystals were characterized by precession photography, and native data sets of each channel-forming fragment were collected on a Siemens-Nicolet area detector. The crystallization and characterization of these polypeptides are the first steps in the structure determination of the channel-forming domain of colicin E1. (C) 1994 Wiley-Liss, Inc.

引用

页码：150 / 157

页数：8

共 30 条

[1] LOCALIZATION OF THE IMMUNITY PROTEIN-REACTIVE DOMAIN IN UNMODIFIED AND CHEMICALLY MODIFIED COOH-TERMINAL PEPTIDES OF COLICIN E1 [J].