Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analysis of the mannose 6-phosphate isomerase from Salmonella typhimurium.

ABSTRACT: Mannose 6-phosphate isomerase (MPI; EC 5.3.1.8) catalyzes the reversible isomerization of D-mannose 6-phosphate (M6P) and D-fructose 6-phosphate (F6P). In the eukaryotes and prokaryotes investigated to date, the enzyme has been reported to play a crucial role in D-mannose metabolism and supply of the activated mannose donor guanosine diphosphate D-mannose (GDP-D-mannose). In the present study, MPI was cloned from Salmonella typhimurium, overexpressed in Escherichia coli and purified using Ni-NTA affinity column chromatography. Purified MPI crystallized in space group P2(1)2(1)2(1), with unit-cell parameters a = 36.03, b = 92.2, c = 111.01 A. A data set extending to 1.66 A resolution was collected with 98.8% completeness using an image-plate detector system mounted on a rotating-anode X-ray

SUBMITTER: Gowda G 

PROVIDER: S-EPMC2374180 | biostudies-literature | 2008 Feb

REPOSITORIES: biostudies-literature