dc.contributor.author | Song, Woon Ju | |
dc.contributor.author | McCormick, Michael S. | |
dc.contributor.author | Behan, Rachel K. | |
dc.contributor.author | Sazinsky, Matthew H. | |
dc.contributor.author | Jiang, Wei | |
dc.contributor.author | Lin, Jeffery | |
dc.contributor.author | Krebs, Carsten | |
dc.contributor.author | Lippard, Stephen J. | |
dc.date.accessioned | 2011-12-14T21:17:05Z | |
dc.date.available | 2011-12-14T21:17:05Z | |
dc.date.issued | 2010-10 | |
dc.date.submitted | 2010-07 | |
dc.identifier.issn | 0002-7863 | |
dc.identifier.issn | 1520-5126 | |
dc.identifier.uri | http://hdl.handle.net/1721.1/67685 | |
dc.description.abstract | Toluene/o-xylene monooxygenase hydroxylase (ToMOH), a diiron-containing enzyme, can activate dioxygen to oxidize aromatic substrates. To elucidate the role of a strictly conserved T201 residue during dioxygen activation of the enzyme, T201S, T201G, T201C, and T201V variants of ToMOH were prepared by site-directed mutagenesis. X-ray crystal structures of all the variants were obtained. Steady-state activity, regiospecificity, and single-turnover yields were also determined for the T201 mutants. Dioxygen activation by the reduced T201 variants was explored by stopped-flow UV−vis and Mossbauer spectroscopy. These studies demonstrate that the dioxygen activation mechanism is preserved in all T201 variants; however, both the formation and decay kinetics of a peroxodiiron(III) intermediate, T201peroxo, were greatly altered, revealing that T201 is critically involved in dioxygen activation. A comparison of the kinetics of O2 activation in the T201S, T201C, and T201G variants under various reaction conditions revealed that T201 plays a major role in proton transfer, which is required to generate the peroxodiiron(III) intermediate. A mechanism is postulated for dioxygen activation, and possible structures of oxygenated intermediates are discussed. | en_US |
dc.description.sponsorship | National Institute of General Medical Sciences (U.S.) (grant GM32134) | en_US |
dc.description.sponsorship | Research Corporation | en_US |
dc.description.sponsorship | National Institute of General Medical Sciences (U.S.) (NIGMS (1 F32 GM084564-02)) | en_US |
dc.language.iso | en_US | |
dc.publisher | American Chemical Society | en_US |
dc.relation.isversionof | http://dx.doi.org/10.1021/ja1063795 | en_US |
dc.rights | Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. | en_US |
dc.source | Prof. Lippard via Erja Kajosalo | en_US |
dc.title | Active Site Threonine Facilitates Proton Transfer during Dioxygen Activation at the Diiron Center of Toluene/o-Xylene Monooxygenase Hydroxylase | en_US |
dc.type | Article | en_US |
dc.identifier.citation | Song, Woon Ju et al. “Active Site Threonine Facilitates Proton Transfer during Dioxygen Activation at the Diiron Center of Toluene/o-xylene Monooxygenase Hydroxylase.” Journal of the American Chemical Society 132.39 (2010): 13582-13585. | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Department of Chemistry | en_US |
dc.contributor.approver | Lippard, Stephen J. | |
dc.contributor.mitauthor | Song, Woon Ju | |
dc.contributor.mitauthor | McCormick, Michael S. | |
dc.contributor.mitauthor | Behan, Rachel K. | |
dc.relation.journal | Journal of the American Chemical Society | en_US |
dc.eprint.version | Author's final manuscript | en_US |
dc.type.uri | http://purl.org/eprint/type/JournalArticle | en_US |
eprint.status | http://purl.org/eprint/status/PeerReviewed | en_US |
dspace.orderedauthors | Song, Woon Ju; McCormick, Michael S.; Behan, Rachel K.; Sazinsky, Matthew H.; Jiang, Wei; Lin, Jeffery; Krebs, Carsten; Lippard, Stephen J. | en |
mit.license | PUBLISHER_POLICY | en_US |
mit.metadata.status | Complete | |