title: Molecular tools for investigating ANME community structure and function.
abstract: Methane production and consumption in anaerobic marine sediments is catalyzed by a series of reversible tetrahydromethanopterin (H(4)MPT)-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.
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Hallam
authors.forename: Antoine P
authors.initials: AP
authors.lastname: Pagé
authors.forename: Lea
authors.initials: L
authors.lastname: Constan
authors.forename: Young C
authors.initials: YC
authors.lastname: Song
authors.forename: Angela D
authors.initials: AD
authors.lastname: Norbeck
authors.forename: Heather
authors.initials: H
authors.lastname: Brewer
authors.forename: Ljiljana
authors.initials: L
authors.lastname: Pasa-Tolic
mesh_terms: D000693:Anaerobiosis; D001105:Archaea / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D019843:Archaeal Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D008697:Methane / Q000096:biosynthesis / Q000378:metabolism*; D010088:Oxidoreductases / Q000145:classification / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D053719:Tandem Mass Spectrometry
chemical_list: D019843:Archaeal Proteins; D010088:Oxidoreductases; C021572:methyl coenzyme M reductase; D008697:Methane