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Molecular Tools for Investigating ANME Community Structure and Function

Authors
  • Hallam, Steven J.
  • P. Pagé, Antoine
  • Constan, Lea
  • Song, Young C.
  • D. Norbeck, Angela
  • Brewer, Heather
  • Pasa-Tolic, Ljiljana
Type
Book
Journal
G Protein Coupled Receptors - Structure
Publisher
Elsevier BV
Publication Date
Jan 01, 2011
Volume
494
Pages
75–90
Identifiers
DOI: 10.3791/1569
ISBN: 978-0-12-385112-3
Source
Elsevier
License
Unknown

Abstract

Methane production and consumption in anaerobic marine sediments is catalyzed by a series of reversible tetrahydromethanopterin (H 4MPT)-linked C1 transfer reactions. Although many of these reactions are conserved between one-carbon compound utilizing microorganisms, two remain diagnostic for archaeal methane metabolism. These include reactions catalyzed by N5-methyltetrahydromethanopterin: coenzyme M methyltransferase and methyl-coenzyme M reductase (MCR). The latter enzyme is central to C–H bond formation and cleavage underlying methanogenic and reverse methanogenic phenotypes. Here, we describe a set of novel tools for the detection and quantification of H4MPT-linked C1 transfer reactions mediated by uncultivated anaerobic methane-oxidizing archaea (ANME). These tools include polymerase chain reaction primers targeting ANME MCR subunit A subgroups and protein extraction methods from marine sediments compatible with high-resolution mass spectrometry for profiling community structure and functional dynamics.

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