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

Gowda, Giri and Sagurthi, Someswar Rao and Savithri, HS and Murthy, MRN (2008) Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analysis of the mannose 6-phosphate isomerase from Salmonella typhimurium. In: Acta Crystallographica Section F Structural Biology and Crystallization Communications, 64 . pp. 81-84.

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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 $P2_12_12_1$, with unit-cell parameters a = 36.03, b = 92.2, c = 111.01 \AA. A data set extending to 1.66 \AA resolution was collected with 98.8% completeness using an image-plate detector system mounted on a rotating-anode X-ray generator. The asymmetric unit of the crystal cell was compatible with the presence of a monomer of MPI. A preliminary structure solution of the enzyme has been obtained by molecular replacement using Candida albicans MPI as the phasing model and the program Phaser. Further refinement and model building are in progress.

Item Type:	Journal Article
Publication:	Acta Crystallographica Section F Structural Biology and Crystallization Communications
Publisher:	International Union of Crystallography
Additional Information:	Copyright of this article belongs to International Union of Crystallography
Keywords:	mannose 6-phosphate isomerase; mannose 6-phosphate; mannose metabolism; Salmonella typhimurium.
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	08 Mar 2008
Last Modified:	19 Sep 2010 04:43
URI:	http://eprints.iisc.ac.in/id/eprint/13357

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