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PDBsum entry 4fdv
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References listed in PDB file
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Key reference
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Title
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An enzyme-Trap approach allows isolation of intermediates in cobalamin biosynthesis.
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Authors
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E.Deery,
S.Schroeder,
A.D.Lawrence,
S.L.Taylor,
A.Seyedarabi,
J.Waterman,
K.S.Wilson,
D.Brown,
M.A.Geeves,
M.J.Howard,
R.W.Pickersgill,
M.J.Warren.
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Ref.
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Nat Chem Biol, 2012,
8,
933-940.
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PubMed id
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Abstract
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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.
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