Finding intermediates in the O2 activation pathways of non-heme iron oxygenases

E. G. Kovaleva, M. B. Neibergall, S. Chakrabarty, John D. Lipscomb

Research output: Contribution to journalArticlepeer-review

178 Scopus citations

Abstract

Intermediates in the reaction cycle of an oxygenase are usually very informative with respect to the chemical mechanism of O2 activation and insertion. However, detection of these intermediates is often complicated by their short lifetime and the regulatory mechanism of the enzyme designed to ensure specificity. Here, the methods used to detect the intermediates in an extradiol dioxygenase, a Rieske cis-dihydrodiol dioxygenase, and soluble methane monooxygenase are discussed. The methods include the use of alternative, chromophoric substrates, mutagenesis of active site catalytic residues, forced changes in substrate binding order, control of reaction rates using regulatory proteins, and initialization of catalysis in crystallo.

Original languageEnglish (US)
Pages (from-to)475-483
Number of pages9
JournalAccounts of Chemical Research
Volume40
Issue number7
DOIs
StatePublished - Jul 2007

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