Metal reconstitution of particulate methane Monooxygenase and heterologous expression of the pmoB subunit

Stephen M. Smith, Ramakrishnan Balasubramanian, Amy C. Rosenzweig

Research output: Chapter in Book/Report/Conference proceedingChapter

26 Scopus citations

Abstract

Particulate methane monooxygenase (pMMO) is a multisubunit metalloenzyme complex used by methanotrophic bacteria to oxidize methane in the first step of carbon assimilation and energy production. In this chapter, we detail methods to prepare metal free (apo) membrane-bound pMMO and to reconstitute apo pMMO with metal ions. We also describe protocols to clone, express, and refold metal-loaded soluble domain constructs of the pmoB subunit. These approaches were used to address fundamental questions concerning the metal content and location of the pMMO active site.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc
Pages195-210
Number of pages16
DOIs
StatePublished - 2011

Publication series

NameMethods in Enzymology
Volume495
ISSN (Print)0076-6879

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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