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Chromophore function and interaction in Escherichia coli DNA photolyase: Reconstitution of the apoenzyme with pterin and/or flavin derivatives
Journal article   Peer reviewed

Chromophore function and interaction in Escherichia coli DNA photolyase: Reconstitution of the apoenzyme with pterin and/or flavin derivatives

M S Jorns, Baoyu Wang, S Jordan and L Chanderkar
Biochemistry (Easton), v 29(2), pp 552-561
01 Jan 1990
PMID: 2405908

Abstract

Escherichia coli
Native DNA photolyase, as isolated from Escherichia coli , contains a neutral flavin radical (FADH) plus a pterin chromophore (5,10-methenyltetrahydropteroyl-polyglutamate) and can be converted to its physiologically significant form by reduction of FADH to fully reduced flavin (FADH sub(2)) with dithionite or by photoreduction. Various enzyme forms (EFAD sub(ox), EFADH, EFADH sub(2), EPteFAD sub(ox), EPteFADH, EPteFADH sub(2), EPte) containing stoichiometric amounts of FAD in either of its three oxidation states and/or 5,10-methenyltetrahydrofolate (Pte) have been prepared in reconstitution experiments. Studies with EFAD sub(ox) and EPte showed that these could be these preparations retained the ability to bind the missing chromophore. The results suggest that there could be considerable flexibility in the biological assembly of holoenzyme since the order of binding of the enzyme's chromophores is apparently unimportant, the binding of FAD is unaffected by its redox state, and enzyme preparations containing only one chromophore are reasonably stable.

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Biochemistry & Molecular Biology
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