Localization of Methyl-Coenzyme M Reductase as Metabolic Marker for Diverse Methanogenic Archaea

Methyl-Coenzyme M reductase (MCR) as key enzyme for methanogenesis as well as for anaerobic oxidation of methane represents an important metabolic marker for both processes in microbial biofilms. Here, the potential of MCR-specific polyclonal antibodies as metabolic marker in various methanogenic Archaea is shown. For standard growth conditions in laboratory culture, the cytoplasmic localization of the enzyme in Methanothermobacter marburgensis, Methanothermobacter wolfei, Methanococcus maripaludis, Methanosarcina mazei, and in anaerobically methane-oxidizing biofilms is demonstrated. Under growth limiting conditions on nickel-depleted media, at low linear growth of cultures, a fraction of 50–70% of the enzyme was localized close to the cytoplasmic membrane, which implies “facultative” membrane association of the enzyme. This feature may be also useful for assessment of growth-limiting conditions in microbial biofilms.

[1]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[2]  R. Thauer,et al.  Differential expression of the two methyl-coenzyme M reductases in Methanobacterium thermoautotrophicum as determined immunochemically via isoenzyme-specific antisera. , 1992, European journal of biochemistry.

[3]  G. Gottschalk,et al.  The Na(+)-translocating methyltransferase complex from methanogenic archaea. , 2001, Biochimica et biophysica acta.

[4]  H. Gross,et al.  Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels , 1987 .

[5]  M. Hoppert,et al.  Cloning, sequencing and immunological characterization of the corrinoid-containing subunit of the N5-methyltetrahydromethanopterin: coenzyme-M methyltransferase from Methanobacterium thermoautotrophicum. , 1993, European journal of biochemistry.

[6]  R. Thauer,et al.  Inhibition of factor F430 synthesis by levulinic acid in Methanobacterium thermoautotrophicum , 1981 .

[7]  R. Thauer,et al.  Growth parameters (Ks, μmax, Ys) of Methanobacterium thermoautotrophicum , 1980, Archives of Microbiology.

[8]  R. Amann,et al.  Metagenome and mRNA expression analyses of anaerobic methanotrophic archaea of the ANME-1 group. , 2010, Environmental microbiology.

[9]  R. Thauer,et al.  Properties of the two isoenzymes of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum. , 1993, European journal of biochemistry.

[10]  H. König,et al.  Methanobacterium wolfei, sp. nov., a New Tungsten-Requiring, Thermophilic, Autotrophic Methanogen , 1984 .

[11]  Daniel Rokhsar,et al.  Reverse Methanogenesis: Testing the Hypothesis with Environmental Genomics , 2004, Science.

[12]  C. Walsh,et al.  Nickel tetrapyrrole cofactor F430: comparison of the forms bound to methyl S-coenzyme M reductase and protein free in cells of Methanobacterium thermoautotrophicum .DELTA.H , 1984 .

[13]  R. Hedderich,et al.  Physiological role of the F420-non-reducing hydrogenase (Mvh) from Methanothermobacter marburgensis , 2003, Archives of Microbiology.

[14]  G. Gottschalk,et al.  Immunocytochemical localization of component C of the methylreductase system in Methanococcus voltae and Methanobacterium thermoautotrophicum. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[15]  V. Thiel,et al.  Concretionary methane-seep carbonates and associated microbial communities in Black Sea sediments , 2005 .

[16]  R. Thauer,et al.  Two genetically distinct methyl-coenzyme M reductases in Methanobacterium thermoautotrophicum strain Marburg and delta H. , 1990, European journal of biochemistry.

[17]  H. Klenk,et al.  Characterization and phylogeny of mcrII, a gene cluster encoding an isoenzyme of methyl coenzyme M reductase from hyperthermophilic Methanothermus fervidus , 1994, Molecular and General Genetics MGG.

[18]  H. Aldrich,et al.  Immunocytochemical localization of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum , 1987, Archives of Microbiology.

[19]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[20]  M. Hoppert,et al.  Principles of macromolecular organization and cell function in bacteria and archaea , 2007, Cell Biochemistry and Biophysics.

[21]  M. Hoppert,et al.  Methods for in situ detection and characterization of extracellular polymers in biofilms by electron microscopy. , 2004, Journal of microbiological methods.

[22]  G. Gottschalk,et al.  The methanoreductosome: a high-molecular-weight enzyme complex in the methanogenic bacterium strain Gö1 that contains components of the methylreductase system , 1988, Journal of bacteriology.

[23]  Anne-Kristin Kaster,et al.  Methanogenic archaea: ecologically relevant differences in energy conservation , 2008, Nature Reviews Microbiology.

[24]  Rudolf Amann,et al.  A conspicuous nickel protein in microbial mats that oxidize methane anaerobically , 2003, Nature.

[25]  P. Debeire,et al.  Purification of component C from Methanosarcina mazei and immunolocalization in Methanosarcinaceae , 1987, Archives of Microbiology.

[26]  H. König,et al.  Methanococcus thermolithotrophicus, a novel thermophilic lithotrophic methanogen , 1982, Archives of Microbiology.

[27]  R. Thauer.  Biochemistry of methanogenesis: a tribute to Marjory Stephenson. 1998 Marjory Stephenson Prize Lecture. , 1998, Microbiology.

[28]  R. Thauer,et al.  Methyl-coenzyme-M reductase from Methanobacterium thermoautotrophicum (strain Marburg). Purity, activity and novel inhibitors. , 1989, European journal of biochemistry.

[29]  L. Daniels,et al.  Dinitrogen fixation by a thermophilic methanogenic bacterium , 1984, Nature.

[30]  R. Fleischmann,et al.  Complete Genome Sequence of the Methanogenic Archaeon, Methanococcus jannaschii , 1996, Science.

[31]  E. Delong,et al.  Methane-Consuming Archaea Revealed by Directly Coupled Isotopic and Phylogenetic Analysis , 2001, Science.

[32]  M. Hoppert,et al.  Electron microscopy of native and artificial methylreductase high‐molecular‐weight complexes in strain Gö 1 and Methanococcus voltae , 1990, FEBS letters.

[33]  M. Hoppert,et al.  Detection of Metabolic Key Enzymes of Methane Turnover Processes in Cold Seep Microbial Biofilms , 2013 .

[34]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[35]  R. Thauer,et al.  The final step in methane formation. Investigations with highly purified methyl-CoM reductase (component C) from Methanobacterium thermoautotrophicum (strain Marburg). , 1988, European journal of biochemistry.

[36]  Martin Krueger,et al.  Structure of a methyl-coenzyme M reductase from Black Sea mats that oxidize methane anaerobically , 2011, Nature.

[37]  U. Deppenmeier.  Redox-driven proton translocation in methanogenic Archaea , 2002, Cellular and Molecular Life Sciences CMLS.

[38]  H. König,et al.  Isolation and characterization of Methanolobus tindarius, sp. nov., a coccoid methanogen growing only on methanol and methylamines , 1982 .

[39]  C. Woese,et al.  Partial gene sequences for the A subunit of methyl-coenzyme M reductase (mcrI) as a phylogenetic tool for the family Methanosarcinaceae. , 1995, International journal of systematic bacteriology.

[40]  Peter G. Brewer,et al.  Methane-consuming archaebacteria in marine sediments , 1999, Nature.

[41]  M. Hoppert,et al.  Immunological Localization of Coenzyme M Reductase in Anaerobic Methane-Oxidizing Archaea of ANME 1 and ANME 2 Type , 2008 .

[42]  K. Fiebig,et al.  Utilization of trimethylamine and other N-methyl compounds for growth and methane formation by Methanosarcina barkeri. , 1979, Proceedings of the National Academy of Sciences of the United States of America.