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Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of propionate kinase (TdcD) from Salmonella typhimurium

Simanshu, Dhirendra K and Murthy, MRN (2005) Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of propionate kinase (TdcD) from Salmonella typhimurium. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 61 (Part 1). pp. 52-55.

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Abstract

In the cell, propionate is mainly formed during \beta-oxidation of odd-numbered carbon-chain fatty acids, fermentation of carbohydrates and degradation of the amino acids threonine, valine, isoleucine and methionine. Recently, it has been shown that L-threonine is non-oxidatively cleaved to propionate via 2-ketobutyrate. The last step in this process, conversion of propionyl phosphate and ADP to propionate and ATP, is catalysed by propionate kinase (EC 2.7.1.-). Here, the cloning of propionate kinase (molecular weight 44 kDa) from Salmonella typhimurium with an N-terminal hexahistidine affinity tag and its overexpression in Escherichia coli are reported. Purified propionate kinase was found to cocrystallize with ADP in the hanging-drop vapour-diffusion and microbatch methods. Crystals belong to space group $P3_{1}21$ or $P3_{2}21$, with unit-cell parameters a = b = 111.47, c = 66.52 Angstron…. A complete data set to 2.2 Angstron… resolution has been collected using an image-plate detector system mounted on a rotating-anode X-ray generator.

Item Type: Journal Article
Additional Information: Copyright for this article belongs to Blackwell Publishing.
Keywords: TdcD;propionate kinases;acetate kinases;L-threonine metabolism
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 24 Oct 2005
Last Modified: 19 Sep 2010 04:20
URI: http://eprints.iisc.ernet.in/id/eprint/3885

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