Deery, Evelyne and Schroeder, Susanne and Lawrence, Andrew D. and Taylor, Samantha L and Seyedarabi, Arefeh and Waterman, Jitka and Wilson, Keith S and Brown, David G. and Geeves, Michael A. and Howard, Mark J. and Pickersgill, Richard W. and Warren, Martin J. (2012) An enzyme-trap approach allows isolation of intermediates in cobalamin biosynthesis. Nature Chemical Biology, 8 (11). pp. 933-940. ISSN 1552-4450. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)
The biosynthesis of many vitamins and coenzymes has often proven difficult to elucidate owing to a combination of low abundance and kinetic lability of the pathway intermediates. Through a serial reconstruction of the cobalamin (vitamin B 12) pathway in Escherichia coli and by His tagging the terminal enzyme in the reaction sequence, we have observed that many unstable intermediates can be isolated as tightly bound enzyme-product complexes. Together, these approaches have been used to extract intermediates between precorrin-4 and hydrogenobyrinic acid in their free acid form and permitted the delineation of the overall reaction catalyzed by CobL, including the formal elucidation of precorrin-7 as a metabolite. Furthermore, a substrate-carrier protein, CobE, that can also be used to stabilize some of the transient metabolic intermediates and enhance their onward transformation, has been identified. The tight association of pathway intermediates with enzymes provides evidence for a form of metabolite channeling.
|Uncontrolled keywords:||article; biosynthesis; catalysis; chemical reaction; complex formation; enzyme activity; enzyme substrate; Escherichia coli; metabolite; nonhuman; priority journal; carrier protein; cobalamin; cobamamide; precorrin; unclassified drug; uroporphyrinogen|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Sue Davies|
|Date Deposited:||09 Oct 2012 10:50|
|Last Modified:||16 Jul 2014 11:35|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/31428 (The current URI for this page, for reference purposes)|