Open AccessCloning, expression and crystallization of dihydrodipicolinate reductase from methicillin‐resistant Staphylococcus aureus
Author(s) -
Dommaraju Sudhir,
Gorman Michael A.,
Dogovski Con,
Pearce F. Grant,
Gerrard Juliet A.,
Dobson Renwick C. J.,
Parker Michael W.,
Perugini Matthew A.
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109047964
Subject(s) - cloning (programming) , cofactor , staphylococcus aureus , enzyme , crystallization , reductase , biosynthesis , ammonium sulfate , escherichia coli , biochemistry , bacteria , biology , stereochemistry , chemistry , microbiology and biotechnology , gene , genetics , organic chemistry , computer science , chromatography , programming language
Dihydrodipicolinate reductase (DHDPR; EC 1.3.1.26) catalyzes the nucleotide (NADH/NADPH) dependent second step of the lysine‐biosynthesis pathway in bacteria and plants. Here, the cloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of DHDPR from methicillin‐resistant Staphylococcus aureus (MRSA‐DHDPR) are presented. The enzyme was crystallized in a number of forms, predominantly with ammonium sulfate as a precipitant, with the best crystal form diffracting to beyond 3.65 Å resolution. Crystal structures of the apo form as well as of cofactor (NADPH) bound and inhibitor (2,6‐pyridinedicarboxylate) bound forms of MRSA‐DHDPR will provide insight into the structure and function of this essential enzyme and valid drug target.