Open AccessPurification, crystallization and preliminary X‐ray diffraction studies to near‐atomic resolution of dihydrodipicolinate synthase from methicillin‐resistant Staphylococcus aureus
Author(s) -
Burgess Benjamin R.,
Dobson Renwick C. J.,
Dogovski Con,
Jameson Geoffrey B.,
Parker Michael W.,
Perugini Matthew A.
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108016746
Subject(s) - lyase , crystallization , staphylococcus aureus , enzyme , chemistry , molecular replacement , pathogen , crystallography , biochemistry , microbiology and biotechnology , stereochemistry , biology , bacteria , organic chemistry , genetics
In recent years, dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) has received considerable attention from both mechanistic and structural viewpoints. DHDPS is part of the diaminopimelate pathway leading to lysine, coupling ( S )‐aspartate‐β‐semialdehyde with pyruvate via a Schiff base to a conserved active‐site lysine. In this paper, the cloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of DHDPS from methicillin‐resistant Staphylococcus aureus , an important bacterial pathogen, are reported. The enzyme was crystallized in a number of forms, predominantly from PEG precipitants, with the best crystal diffracting to beyond 1.45 Å resolution. The space group was P 1 and the unit‐cell parameters were a = 65.4, b = 67.6, c = 78.0 Å, α = 90.1, β = 68.9, γ = 72.3°. The crystal volume per protein weight ( V M ) was 2.34 Å 3 Da −1 , with an estimated solvent content of 47% for four monomers per asymmetric unit. The structure of the enzyme will help to guide the design of novel therapeutics against the methicillin‐resistant S. aureus pathogen.