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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Wang, Xiaoshi Ullrich, René Hofrichter, Martin Groves, John T. |
| Description | Author Affiliation: Wang X ( Department of Chemistry, Princeton University, Princeton, NJ 08544); Ullrich R ( Department of Bio- and Environmental Sciences, International Institute Zittau, Dresden University of Technology, 02763 Zittau, Germany.); Hofrichter M ( Department of Bio- and Environmental Sciences, International Institute Zittau, Dresden University of Technology, 02763 Zittau, Germany.); Groves JT ( Department of Chemistry, Princeton University, Princeton, NJ 08544); |
| Abstract | A kinetic and spectroscopic characterization of the ferryl intermediate (APO-II) from APO, the heme-thiolate peroxygenase from Agrocybe aegerita, is described. APO-II was generated by reaction of the ferric enzyme with metachloroperoxybenzoic acid in the presence of nitroxyl radicals and detected with the use of rapid-mixing stopped-flow UV-visible (UV-vis) spectroscopy. The nitroxyl radicals served as selective reductants of APO-I, reacting only slowly with APO-II. APO-II displayed a split Soret UV-vis spectrum (370 nm and 428 nm) characteristic of thiolate ligation. Rapid-mixing, pH-jump spectrophotometry revealed a basic pKa of 10.0 for the Fe(IV)-O-H of APO-II, indicating that APO-II is protonated under typical turnover conditions. Kinetic characterization showed that APO-II is unusually reactive toward a panel of benzylic C-H and phenolic substrates, with second-order rate constants for C-H and O-H bond scission in the range of 10-10(7) M(-1)â s(-1). Our results demonstrate the important role of the axial cysteine ligand in increasing the proton affinity of the ferryl oxygen of APO intermediates, thus providing additional driving force for C-H and O-H bond scission. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 12 |
| Volume Number | 112 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2015-03-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Agrocybe Enzymology Heme Chemistry Mixed Function Oxygenases Sulfhydryl Compounds Hydrogen Hydrogen-Ion Concentration Nitrogen Oxidation-Reduction Oxygen Phenol Spectrophotometry, Ultraviolet Substrate Specificity Temperature Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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