Open AccessCloning, expression, purification and crystallization of dihydrodipicolinate synthase from the psychrophile Shewanella benthica
Author(s) -
Wubben Jacinta M.,
Dogovski Con,
Dobson Renwick C. J.,
Codd Rachel,
Gerrard Juliet A.,
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/s1744309110036791
Subject(s) - psychrophile , protein quaternary structure , thermophile , enzyme , shewanella , lyase , biology , biochemistry , bacteria , chemistry , protein subunit , gene , genetics
Dihydrodipicolinate synthase (DHDPS) is an oligomeric enzyme that catalyzes the first committed step of the lysine‐biosynthesis pathway in plants and bacteria, which yields essential building blocks for cell‐wall and protein synthesis. DHDPS is therefore of interest to drug‐discovery research as well as to studies that probe the importance of quaternary structure to protein function, stability and dynamics. Accordingly, DHDPS from the psychrophilic (cold‐dwelling) organism Shewanella benthica ( Sb ‐DHDPS) was cloned, expressed, purified and crystallized. The best crystals of Sb ‐DHDPS were grown in 200 m M ammonium sulfate, 100 m M bis‐tris pH 5.0–6.0, 23–26%( w / v ) PEG 3350, 0.02%( w / v ) sodium azide and diffracted to beyond 2.5 Å resolution. Processing of diffraction data to 2.5 Å resolution resulted in a unit cell with space group P 2 1 2 1 2 1 and dimensions a = 73.1, b = 84.0, c = 143.7 Å. These studies of the first DHDPS enzyme to be characterized from a bacterial psychrophile will provide insight into the molecular evolution of enzyme structure and dynamics.