Evaluation of a concerted vs. sequential oxygen activation mechanism in α-ketoglutarate–dependent nonheme ferrous enzymes

Goudarzi, Serra; Iyer, Shyam R.; Babicz, Jr., Jeffrey T.; Yan, James J.; Peters, Günther H.  J.; Christensen, Hans E.  M.; Hedman, Britt; Hodgson, Keith O.; Solomon, Edward I.

doi:10.1073/pnas.1922484117

Evaluation of a concerted vs. sequential oxygen activation mechanism in α-ketoglutarate–dependent nonheme ferrous enzymes

Journal Article · Mon Feb 24 00:00:00 EST 2020 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1922484117· OSTI ID:1608322

Goudarzi, Serra ^[1]; Iyer, Shyam R. ^[1]; Babicz, Jr., Jeffrey T. ^[1]; ^[2]; ^[3]; Christensen, Hans E. M. ^[3]; Hedman, Britt ^[4]; Hodgson, Keith O. ^[2]; ^[2]

Stanford Univ., CA (United States)
Stanford Univ., CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)
Technical Univ. of Denmark, Lyngby (Denmark)
SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Determining the requirements for efficient oxygen (O₂) activation is key to understanding how enzymes maintain efficacy and mitigate unproductive, often detrimental reactivity. For the α-ketoglutarate (αKG)–dependent nonheme iron enzymes, both a concerted mechanism (both cofactor and substrate binding prior to reaction with O₂) and a sequential mechanism (cofactor binding and reaction with O₂precede substrate binding) have been proposed. Deacetoxycephalosporin C synthase (DAOCS) is an αKG-dependent nonheme iron enzyme for which both of these mechanisms have been invoked to generate an intermediate that catalyzes oxidative ring expansion of penicillin substrates in cephalosporin biosynthesis. Interestingly, spectroscopy shows that, in contrast to other αKG-dependent enzymes (which are six coordinate when only αKG is bound to the Fe^II), αKG binding to Fe^II-DAOCS results in ~45% five-coordinate sites that selectively react with O₂relative to the remaining six-coordinate sites. However, this reaction produces an Fe^IIIspecies that does not catalyze productive ring expansion. Alternatively, simultaneous αKG and substrate binding to Fe^II-DAOCS produces five-coordinate sites that rapidly react with O₂to form an Fe^IV=O intermediate that then reacts with substrate to produce cephalosporin product. These results demonstrate that the concerted mechanism is operative in DAOCS and by extension, other nonheme iron enzymes.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory, Menlo Park, CA (United States)

Sponsoring Organization:: Independent Research Fund Denmark; National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1608322

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 10 Vol. 117; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

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Evaluation of a concerted vs. sequential oxygen activation mechanism in α-ketoglutarate–dependent nonheme ferrous enzymes

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