Diradical intermediate within the context of tryptophan tryptophylquinone biosynthesis.

Abstract:

:Despite the importance of tryptophan (Trp) radicals in biology, very few radicals have been trapped and characterized in a physiologically meaningful context. Here we demonstrate that the diheme enzyme MauG uses Trp radical chemistry to catalyze formation of a Trp-derived tryptophan tryptophylquinone cofactor on its substrate protein, premethylamine dehydrogenase. The unusual six-electron oxidation that results in tryptophan tryptophylquinone formation occurs in three discrete two-electron catalytic steps. Here the exact order of these oxidation steps in the processive six-electron biosynthetic reaction is determined, and reaction intermediates are structurally characterized. The intermediates observed in crystal structures are also verified in solution using mass spectrometry. Furthermore, an unprecedented Trp-derived diradical species on premethylamine dehydrogenase, which is an intermediate in the first two-electron step, is characterized using high-frequency and -field electron paramagnetic resonance spectroscopy and UV-visible absorbance spectroscopy. This work defines a unique mechanism for radical-mediated catalysis of a protein substrate, and has broad implications in the areas of applied biocatalysis and understanding of oxidative protein modification during oxidative stress.

authors

Yukl ET,Liu F,Krzystek J,Shin S,Jensen LM,Davidson VL,Wilmot CM,Liu A

doi

10.1073/pnas.1215011110

subject

Has Abstract

pub_date

2013-03-19 00:00:00

pages

4569-73

issue

12

eissn

0027-8424

issn

1091-6490

pii

1215011110

journal_volume

110

pub_type

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