Raux, E. and Warren, M.J. (1998) Cobalamin (vitamin B-12) biosynthesis - Cloning, expression and crystallisation of the Bacillus megaterium S-adenosyl-L-methionine-dependent cobalt-precorrin-4 transmethylase CbiF. European Journal of Biochemistry, 254 (2). pp. 341-346. ISSN 0014-2956 .
|The full text of this publication is not available from this repository. (Contact us about this Publication)|
The Bacillus megaterium cbiF, encoding the cobalt-precorrin-4 S-adenosyl-L-methionine-dependent transmethylase of the anaerobic cobalamin biosynthetic pathway, has been cloned and overexpressed as a His-tagged recombinant protein in Escherichia coli. The protein was purified to homogeneity by a combination of metal chelate chromatography and high-resolution anion-exchange chromatography. The protein migrated with a subunit mass of 31 kDa by SDS/PAGE and with a molecular mass of 62 kDa by analytical gel filtration, suggesting that the native recombinant protein is a homodimer. The His-tagged protein was physiologically active as it was able to complement a Salmonella typhimurium cbiF mutant. However, the protein did not bind S-adenosyl-L-methionine with the same avidity as observed with other corrin biosynthetic transmethylases. A crystallisation screen of the purified protein led to the identification of two discrete crystal forms. One of these forms has been characterised and a full data set collected.
|Uncontrolled keywords:||vitamin b-12 cbif cobalt-preconin-4 bacillus megaterium s-adenosyl-l-methionine uroporphyrinogen-iii methyltransferase pseudomonas-denitrificans salmonella-typhimurium sequence-analysis genes purification synthase intermediate precorrin-2 acid|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Martin Warren|
|Date Deposited:||03 Oct 2009 11:42|
|Last Modified:||14 Jan 2010 14:42|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/11083 (The current URI for this page, for reference purposes)|
- Depositors only (login required):