Open AccessPurification, crystallization and preliminary crystallographic analysis of the adhesion domain of Epf from Streptococcus pyogenes
Author(s) -
Linke Christian,
Siemens Nikolai,
Middleditch Martin J.,
Kreikemeyer Bernd,
Baker Edward N.
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112020313
Subject(s) - streptococcus pyogenes , crystallization , escherichia coli , adhesion , protease , chemistry , extracellular , microbiology and biotechnology , biology , crystallography , biochemistry , bacteria , genetics , enzyme , gene , staphylococcus aureus , organic chemistry
The extracellular protein Epf from Streptococcus pyogenes is important for streptococcal adhesion to human epithelial cells. However, Epf has no sequence identity to any protein of known structure or function. Thus, several predicted domains of the 205 kDa protein Epf were cloned separately and expressed in Escherichia coli . The N‐terminal domain of Epf was crystallized in space groups P 2 1 and P 2 1 2 1 2 1 in the presence of the protease chymotrypsin. Mass spectrometry showed that the species crystallized corresponded to a fragment comprising residues 52–357 of Epf. Complete data sets were collected to 2.0 and 1.6 Å resolution, respectively, at the Australian Synchrotron.