Issue 3, 2009
From the journal:

Organic & Biomolecular Chemistry

Synthesis and use of isotope-labelled substrates for a mechanistic study on human α-methylacyl-CoA racemase 1A (AMACR; P504S)†‡

Daniel J. Darley,a   Danica S. Butler,b   Samuel J. Prideaux,a   Thomas W. Thornton,a   Abigail D. Wilson,a   Timothy J. Woodman,a   Michael D. Threadgilla  and  Matthew D. Lloyd*a  

* Corresponding authors

a Department of Pharmacy & Pharmacology, University of Bath, Claverton Down, Bath, United Kingdom
E-mail: M.D.Lloyd@bath.ac.uk

b Chemistry Research Laboratory, Mansfield Road, University of Oxford, Oxford, United Kingdom

Abstract

α-Methylacyl-CoA racemase (AMACR) is an important enzyme for the metabolism of branched-chain lipids and drugs. The enzyme is over-expressed in prostate and other cancers. AMACR 1A, the major splice variant, was purified from recombinant E. colicells as a His-tag protein. Purified enzyme catalysed chiral inversion of both S- and R-2-methyldecanoyl-CoA, with an equilibrium constant of 1.09 ± 0.14 (2S/2R). Reactions with 2H-labelled substrate showed that loss of the α-proton was a prerequisite for chiral inversion. Reactions conducted in 2H2O indicated that reprotonation was not stereospecific. These results are the first mechanistic study on any recombinant mammalian α-methylacyl-CoA racemase.

Graphical abstract: Synthesis and use of isotope-labelled substrates for a mechanistic study on human α-methylacyl-CoA racemase 1A (AMACR; P504S)

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DOI
https://doi.org/10.1039/B815396E
Article type
Paper
Submitted
03 Sep 2008
Accepted
27 Oct 2008
First published
28 Nov 2008

Org. Biomol. Chem., 2009,7, 543-552

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Synthesis and use of isotope-labelled substrates for a mechanistic study on human α-methylacyl-CoA racemase 1A (AMACR; P504S)

D. J. Darley, D. S. Butler, S. J. Prideaux, T. W. Thornton, A. D. Wilson, T. J. Woodman, M. D. Threadgill and M. D. Lloyd, Org. Biomol. Chem., 2009, 7, 543 DOI: 10.1039/B815396E

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