期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2012
卷号：109
期号：1
页码：39-43
DOI：10.1073/pnas.1115778108
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Photochemical radical initiation is a powerful tool for studying radical initiation and transport in biology. Ribonucleotide reductases (RNRs), which catalyze the conversion of nucleotides to deoxynucleotides in all organisms, are an exemplar of radical mediated transformations in biology. Class Ia RNRs are composed of two subunits: 2 and {beta}2. As a method to initiate radical formation photochemically within {beta}2, a single surface-exposed cysteine of the {beta}2 subunit of Escherichia coli Class Ia RNR has been labeled (98%) with a photooxidant ([Re ] = tricarbonyl(1,10-phenanthroline)(methylpyridyl)rhenium(I)). The labeling was achieved by incubation of S355C-{beta}2 with the 4-(bromomethyl)pyridyl derivative of [Re] to yield the labeled species, [Re]-S355C-{beta}2. Steady-state and time-resolved emission experiments reveal that the metal-to-ligand charge transfer (MLCT) excited-state 3[Re ]* is not significantly perturbed after bioconjugation and is available as a phototrigger of tyrosine radical at position 356 in the {beta}2 subunit; transient absorption spectroscopy reveals that the radical lives for microseconds. The work described herein provides a platform for photochemical radical initiation and study of proton-coupled electron transfer (PCET) in the {beta}2 subunit of RNR, from which radical initiation and transport for this enzyme originates.
关键词：proton-coupled electron transfer ; radical generation ; radical transport
