Open AccessCloning, expression, purification and crystallization of dihydrodipicolinate synthase from the grapevine Vitis vinifera
Author(s) -
Atkinson Sarah C.,
Dogovski Con,
Newman Janet,
Dobson Renwick C. J.,
Perugini Matthew A.
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111038395
Subject(s) - lyase , crystallization , protein quaternary structure , chemistry , stereochemistry , enzyme , biochemistry , organic chemistry , gene , protein subunit
Dihydrodipicolinate synthase (DHDPS) catalyses the first committed step of the lysine‐biosynthesis pathway in bacteria, plants and some fungi. This study describes the cloning, expression, purification and crystallization of DHDPS from the grapevine Vitis vinifera (Vv‐DHDPS). Following in‐drop cleavage of the hexahistidine tag, cocrystals of Vv‐DHDPS with the substrate pyruvate were grown in 0.1 M Bis‐Tris propane pH 8.2, 0.2 M sodium bromide, 20%( w / v ) PEG 3350. X‐ray diffraction data in space group P 1 at a resolution of 2.2 Å are presented. Preliminary diffraction data analysis indicated the presence of eight molecules per asymmetric unit ( V M = 2.55 Å 3 Da −1 , 52% solvent content). The pending crystal structure of Vv‐DHDPS will provide insight into the molecular evolution in quaternary structure of DHDPS enzymes.