Structural aspects and immunolocalization of the F420-reducing and non-F420-reducing hydrogenases from Methanobacterium thermoautotrophicum Marburg
10.1128/jb.176.24.7677-7687.1994
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The F420-reducing hydrogenase and the non-F420-reducing hydrogenase (EC 1.12.99.1.) were isolated from a crude extract of Methanobacterium thermoautotrophicum Marburg. Electron microscopy of the negatively stained F420-reducing hydrogenase revealed that the enzyme is a complex with a diameter of 15.6 nm. It consists of two ring-like, stacked, parallel layers each composed of three major protein masses arranged in rotational symmetry. Each of these masses appeared to be subdivided into smaller protein masses. Electron microscopy of negatively stained samples taken from intermediate steps of the purification process revealed the presence of enzyme particles bound to inside-out membrane vesicles. Linker particles of 10 to 20 kDa which mediate the attachment of the hydrogenase to the cytoplasmic membrane were seen. Immunogold labelling confirmed that the F420-reducing hydrogenase is a membrane-bound enzyme. Electron microscopy of the negatively stained purified non-F420-reducing hydrogenase revealed that the enzyme is composed of three subunits exhibiting different diameters (5, 4, and 2 to 3 nm). According to immunogold labelling experiments, approximately 70% of the non-F420-reducing hydrogenase protein molecules were located at the cell periphery; the remaining 30% were cytoplasmic. No linker particles were observed for this enzyme.

Bibliography

Braks, I. J., Hoppert, M., Roge, S., & Mayer, F. (1994). Structural aspects and immunolocalization of the F420-reducing and non-F420-reducing hydrogenases from Methanobacterium thermoautotrophicum Marburg. Journal of Bacteriology, 176(24), 7677–7687.

Authors 4
  1. I J Braks (first)
  2. M Hoppert (additional)
  3. S Roge (additional)
  4. F Mayer (additional)
References 49 Referenced 23
  1. Aldrich , H. C. , D. B. Beimborn , M. Bokranz , and P. Schonheit . 1987 . Immunocytochemical localization of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum . Arch. Microbiol. 147 : 190 - 194 . (10.1007/BF00415283) / Arch. Microbiol. / Immunocytochemical localization of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum by Aldrich H. C. (1987)
  2. Alex , L A . , J. N. Reeve , W. H. Orme-Johnson , and C. T. Walsh . 1990 . Cloning, sequence determination, and expression of the genes encoding subunits of the Ni containing 8-hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thernoautotrophicum AH . Biochemistry 29 : 7237 - 7244 . (10.1021/bi00483a011) / Biochemistry / Cloning, sequence determination, and expression of the genes encoding subunits of the Ni containing 8-hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thernoautotrophicum AH by Alex L A (1990)
  3. Baron , S. F. , and J. G. Ferry . 1989 . Purification and properties of the membrane-associated coenzyme F420-reducing hydrogenase from Methanobacterium formicicum . J. Bacteriol. 171 : 3846 - 3853 . (10.1128/jb.171.7.3846-3853.1989) / J. Bacteriol. / Purification and properties of the membrane-associated coenzyme F420-reducing hydrogenase from Methanobacterium formicicum by Baron S. F. (1989)
  4. Bates , K. L , M. J. Danson , G. Hale , E. A. Hooper , and R. N. Perham . 1977 . Self assembly and catalytic activity of the pyruvate dehydrogenase multienzyme complex of Escherichia coli . Nature (London) 268 : 313 - 316 . (10.1038/268313a0) / Nature (London) / Self assembly and catalytic activity of the pyruvate dehydrogenase multienzyme complex of Escherichia coli by Bates K. L (1977)
  5. Bauer , R. 1988 . Electron spectroscopic imaging: an advanced technique for imaging and analysis in transmission electron microscopy . Methods Microbiol. 20 : 113 - 146 . (10.1016/S0580-9517(08)70050-3) / Methods Microbiol. / Electron spectroscopic imaging: an advanced technique for imaging and analysis in transmission electron microscopy by Bauer R. (1988)
  6. Blum , H. , H. Beier , and G. J. Gross . 1987 . Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels . Electrophoresis 8 : 93 - 99 . (10.1002/elps.1150080203) / Electrophoresis / Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels by Blum H. (1987)
  7. Deppenmeier , U. , M. Blaut , and G. Gottschalk 1991 . H2-heterodisulfide oxidoreductase, a second energy-conserving system in the methanogenic strain Gol . Arch. Microbiol. 155 : 272 - 277 . (10.1007/BF00252211) / Arch. Microbiol. / H2-heterodisulfide oxidoreductase, a second energy-conserving system in the methanogenic strain Gol by Deppenmeier U. (1991)
  8. Deppenmeier U. M. Blaut C. Herzberg and G. Gottschalk The mvhGAC operon of Methanosarcina mazei strain Got encodes a membrane-bound hydrogenase and a cytochrome b. Submitted for publication.
  9. Fauque , G. , M. Teixeira , I. Moura , P. A. Lespinat , D. V. Der-Vartanian , H. D. Peck , J. Le Gall , and J. G. Moura . 1984 . Purification, characterization and redox properties of hydrogenase from Methanosarcina barkeri (DSM 800) . Eur. J. Biochem. 142 : 21 - 28 . (10.1111/j.1432-1033.1984.tb08245.x) / Eur. J. Biochem. / Purification, characterization and redox properties of hydrogenase from Methanosarcina barkeri (DSM 800) by Fauque G. (1984)
  10. Fiebig , K. , and B. Friedrich . 1989 . Purification of the F420- reducing hydrogenase from Methanosarcina barkeri (strain Fusaro) . Eur. J. Biochem. 184 : 79 - 88 . (10.1111/j.1432-1033.1989.tb14992.x) / Eur. J. Biochem. / Purification of the F420- reducing hydrogenase from Methanosarcina barkeri (strain Fusaro) by Fiebig K. (1989)
  11. Fischer , R. , P. Gairtner , A. Yeliseev , and R. K. Thauer . 1992 . N5-methyl H4MPT: coenzyme M methyltransferase in methanogenic archaebacteria is a membrane protein . Arch. Microbiol. 158 : 208 - 217 . (10.1007/BF00290817) / Arch. Microbiol. / N5-methyl H4MPT: coenzyme M methyltransferase in methanogenic archaebacteria is a membrane protein by Fischer R. (1992)
  12. Fox , J. A. , D. J. Livingston , W. H. Orme-Johnson , and C. T. Walsh . 1987 . 8-Hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum: purification and characterization . Biochemistry 26 : 4219 - 4227 . (10.1021/bi00388a007) / Biochemistry / 8-Hydroxy-5-deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum: purification and characterization by Fox J. A. (1987)
  13. Fuchs G. J. Moll P. Scherer and R. Thauer. 1978. Activity acceptor specification and function of hydrogenase in Methanobacterium thermoautotrophicum p. 83-92. In H. G. Schlegel and K Schneider (ed.) Hydrogenases: their catalytic activity structure and function. Erich Goltze KG Gottingen Germany.
  14. Fuchs , G. , E. Stupperich , and R. K. Thauer . 1978 . Acetate assimilation and the synthesis of alanine, aspartate and glutamate in Methanobacterium thermoautotrophicum . Arch. Microbiol. 117 : 61 - 66 . (10.1007/BF00689352) / Arch. Microbiol. / Acetate assimilation and the synthesis of alanine, aspartate and glutamate in Methanobacterium thermoautotrophicum by Fuchs G. (1978)
  15. Gerberding , H. , and F. Mayer . 1989 . Structural organization of the membrane-bound hydrogenase isolated fromAlcaligenes eutrophus as revealed by electron microscopy . FEMS Microbiol. Lett. 60 : 159 - 164 . (10.1111/j.1574-6968.1989.tb03437.x) / FEMS Microbiol. Lett. / Structural organization of the membrane-bound hydrogenase isolated fromAlcaligenes eutrophus as revealed by electron microscopy by Gerberding H. (1989)
  16. Graf , E. G. , and R. K. Thauer . 1981 . Hydrogenase from Methanobacterium thermoautotrophicum, a nickel-containing enzyme . FEBS Lett. 136 : 165 - 169 . (10.1016/0014-5793(81)81238-0) / FEBS Lett. / Hydrogenase from Methanobacterium thermoautotrophicum, a nickel-containing enzyme by Graf E. G. (1981)
  17. Haase , P. , U. Deppenmeier , M. Blaut , and G. GottschalLk 1992 . Purification and characterization of F420H2-dehydrogenase from Methanolobus tindarius . Eur. J. Biochem. 203 : 527 - 531 . (10.1111/j.1432-1033.1992.tb16579.x) / Eur. J. Biochem. / Purification and characterization of F420H2-dehydrogenase from Methanolobus tindarius by Haase P. (1992)
  18. Hedderich , R. , S. P. J. Albracht , D. Linder , J. Koch , and R. K. Thauer . 1992 . Isolation and characterization of polyferredoxin from Methanobacterium thermoautotrophicum . FEBS Lett. 298 : 65 - 68 . (10.1016/0014-5793(92)80023-A) / FEBS Lett. / Isolation and characterization of polyferredoxin from Methanobacterium thermoautotrophicum by Hedderich R. (1992)
  19. Heiden , S. , R Hedderich , E. Setzke , and R. K. Thauer . 1993 . Purification of a cytochrome-b-containing H2:heterodisulfide oxidoreductase complex from membranes of Methanosarcina barkeri . Eur. J. Biochem. 213 : 529 - 535 . (10.1111/j.1432-1033.1993.tb17791.x) / Eur. J. Biochem. / Purification of a cytochrome-b-containing H2:heterodisulfide oxidoreductase complex from membranes of Methanosarcina barkeri by Heiden S. (1993)
  20. Hurn , B. A. L. , and S. M. Chantler . 1980 . Production of reagent antibodies . Methods Enzymol. 70 : 104 - 142 . (10.1016/S0076-6879(80)70044-7) / Methods Enzymol. / Production of reagent antibodies by Hurn B. A. L. (1980)
  21. Jin , S. L. C. , D. K. Blanchard , and J. S. Chen . 1983 . Two hydrogenases with distinct electron-carrier specificity and subunit composition in Methanobacterium formicicum . Biochim. Biophys. Acta 748 : 8 - 20 . (10.1016/0167-4838(83)90021-3) / Biochim. Biophys. Acta / Two hydrogenases with distinct electron-carrier specificity and subunit composition in Methanobacterium formicicum by Jin S. L. C. (1983)
  22. Jones A. (Ohio State University). 1994. Personal communication.
  23. Jovin , T. , A. Chrambach , and M. A. Naughton . 1964 . An apparatus for temperature-regulated polyacrylamide gel electrophoresis . Anal. Biochem. 9 : 351 - 369 . (10.1016/0003-2697(64)90192-7) / Anal. Biochem. / An apparatus for temperature-regulated polyacrylamide gel electrophoresis by Jovin T. (1964)
  24. Kohring , G. W. , H. Mayer , and F. Mayer . 1985 . Immunoelectron microscopic localisation of the restriction endonuclease EcoRi in Escherichia coli . Eur. J. Cell Biol. 37 : 1 - 6 . / Eur. J. Cell Biol. / Immunoelectron microscopic localisation of the restriction endonuclease EcoRi in Escherichia coli by Kohring G. W. (1985)
  25. Kojima , N. , J. A. Fox , R P . Hausinger , L. Daniels , W. H. Orme-Johnson , and C. T. Walsh . 1983 . Paramagnetic centers in the nickel-containing deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum . Proc. Natl. Acad. Sci. USA 80 . 378 - 382 . (10.1073/pnas.80.2.378) / Proc. Natl. Acad. Sci. USA / Paramagnetic centers in the nickel-containing deazaflavin-reducing hydrogenase from Methanobacterium thermoautotrophicum by Kojima N. (1983)
  26. Laemmli , U. K. 1970 . Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature (London) 227 : 680 - 685 . (10.1038/227680a0) / Nature (London) / Cleavage of structural proteins during the assembly of the head of bacteriophage T4 by Laemmli U. K. (1970)
  27. Lunsdorf , H. , M. Niedrig , and K. Fiebig . 1991 . Immunocytochemical localization of the coenzyme F420-reducing hydrogenase in Methanosarcina barkeri Fusaro . J. Bacteriol. 173 : 1 - 7 . (10.1128/jb.173.3.978-984.1991) / J. Bacteriol. / Immunocytochemical localization of the coenzyme F420-reducing hydrogenase in Methanosarcina barkeri Fusaro by Lunsdorf H. (1991)
  28. Mayer , F. , A. Jussofie , M. Salzmann , M. Lflbben , M. Rohde , and G. Gottschalk . 1987 . Immunoelectron microscopic demonstration of ATPase on the cytoplasmic membrane of the methanogenic bacterium strain Gol . J. Bacteriol. 169 : 2307 - 2309 . (10.1128/jb.169.5.2307-2309.1987) / J. Bacteriol. / Immunoelectron microscopic demonstration of ATPase on the cytoplasmic membrane of the methanogenic bacterium strain Gol by Mayer F. (1987)
  29. McKellar , R. C. , and G. D. Sprott . 1979 . Solubilization and properties of a particulate hydrogenase from Methanobacterium strain G2R . J. Bacteriol. 139 : 231 - 238 . (10.1128/jb.139.1.231-238.1979) / J. Bacteriol. / Solubilization and properties of a particulate hydrogenase from Methanobacterium strain G2R by McKellar R. C. (1979)
  30. Menon , N. K. , J. Robbins , H. D. Peck , Jr. , C. Y. Chatelus , E. Choi , and A. E. Przybyla . 1990 . Cloning and sequencing of a putative Escherichia coli [NiFe] hydrogenase-1 operon containing six open reading frames . J. Bacteriol. 172 : 1969 - 1977 . (10.1128/jb.172.4.1969-1977.1990) / J. Bacteriol. / Cloning and sequencing of a putative Escherichia coli [NiFe] hydrogenase-1 operon containing six open reading frames by Menon N. K. (1990)
  31. Muller V. M. Blaut and G. Gottschalk. 1993. Bioenergetics of methanogenesis p. 360-406. In J. G. Ferry (ed.) Methanogenesis. Chapman & Hall New York. (10.1007/978-1-4615-2391-8_9)
  32. Muth , E. 1987 . Purification and characterization of an 8-hydroxy-5-deazaflavin-reducing hydrogenase from the archaebacterium Methanococcus voltae . Eur. J. Biochem. 169 : 571 - 577 . (10.1111/j.1432-1033.1987.tb13647.x) / Eur. J. Biochem. / Purification and characterization of an 8-hydroxy-5-deazaflavin-reducing hydrogenase from the archaebacterium Methanococcus voltae by Muth E. (1987)
  33. Ossmer , R. , T. Mund , P. L. Hartzell , U. Konheiser , G. W. Kohring , A. Klein , R S . Wolfe , G. Gottschalk , and F. Mayer . 1986 . Immunocytochemical localization of component C of the methylreductase system in Methanococcus voltae and Methanobacterium thermoautotrophicum . Proc. Natl. Acad. Sci. USA 83 : 5789 - 5792 . (10.1073/pnas.83.16.5789) / Proc. Natl. Acad. Sci. USA / Immunocytochemical localization of component C of the methylreductase system in Methanococcus voltae and Methanobacterium thermoautotrophicum by Ossmer R. (1986)
  34. Ottensmeyer , F. P. , and J. W. Andrew . 1980 . High-resolution microanalysis of biological specimens by electron energy loss spectroscopy and by electron spectroscopic imaging . J. Ultrastruct. Res. 72 : 336 - 348 . (10.1016/S0022-5320(80)90069-6) / J. Ultrastruct. Res. / High-resolution microanalysis of biological specimens by electron energy loss spectroscopy and by electron spectroscopic imaging by Ottensmeyer F. P. (1980)
  35. Przybyla , A. E. , J. Robbins , N. Menon , and H. D. Peck , Jr. 1992 . Structure-function relationships among the nickel-containing hydrogenases . FEMS Microbiol. Rev. 88 : 109 - 136 . (10.1111/j.1574-6968.1992.tb04960.x) / FEMS Microbiol. Rev. / Structure-function relationships among the nickel-containing hydrogenases by Przybyla A. E. (1992)
  36. Reeve , J. N. , and G. S. Beckler . 1990 . Conservation of primary structure in prokaryotic hydrogenases . FEMS Microbiol. Rev. 87 : 419 - 424 . (10.1111/j.1574-6968.1990.tb04947.x) / FEMS Microbiol. Rev. / Conservation of primary structure in prokaryotic hydrogenases by Reeve J. N. (1990)
  37. Reeve , J. N. , G. S. Beckler , D. S. Cram , P. T. Hamilton , J. W. Brown , J. A. Krzycki , A. F. Kolodziej , L. Alex , W. H. Orme-Johnson , and C. T. Walsh . 1989 . A hydrogenase-linked gene in Methanobactenium thermoautotrophicum strain AH encodes a polyferredoxin . Proc. Natl. Acad. Sci. USA 86 : 3031 - 3035 . (10.1073/pnas.86.9.3031) / Proc. Natl. Acad. Sci. USA / A hydrogenase-linked gene in Methanobactenium thermoautotrophicum strain AH encodes a polyferredoxin by Reeve J. N. (1989)
  38. Reimer L. 1991. Energy-filtering transmission electron microscopy p. 41-126. In P. W. Hawkes (ed.) Advances in electronics and electron physics vol. 81. Academic Press Inc. New York. (10.1016/S0065-2539(08)60863-X)
  39. Rohde , M. , H. Gerberding , T. Mund , and G. W. Kohring . 1988 . Immunoelectron microscopic localisation of bacterial enzymes: pre- and post-embedding labelling techniques on resin-embedded samples . Methods Microbiol. 20 : 175 - 210 . (10.1016/S0580-9517(08)70052-7) / Methods Microbiol. / Immunoelectron microscopic localisation of bacterial enzymes: pre- and post-embedding labelling techniques on resin-embedded samples by Rohde M. (1988)
  40. Schonheit , P. , J. Moll , and R. K. Thauer . 1980 . Growth parameters (Ks, mmax, Ys) of Methanobacterium thermoautotrophicum . Arch. Microbiol. 127 : 59 - 65 . (10.1007/BF00414356) / Arch. Microbiol. / Growth parameters (Ks, mmax, Ys) of Methanobacterium thermoautotrophicum by Schonheit P. (1980)
  41. Setzke , E. , R Hedderich , S. Heiden , and R K . Thauer . 1994 . H2:heterodisulfide oxidoreductase complex from Methanobactenium thermoautotrophicum. Composition and properties . Eur. J. Biochem. 220 : 139 - 148 . (10.1111/j.1432-1033.1994.tb18608.x) / Eur. J. Biochem. / H2:heterodisulfide oxidoreductase complex from Methanobactenium thermoautotrophicum. Composition and properties by Setzke E. (1994)
  42. Shermann , M. B. , E. V. Orlova , E. A. Smirnova , S. Hovmoller , and N. A. Zorin . 1991 . Three-dimensional structure of the nickelcontaining hydrogenase from Thiocapsa roseopersicina . J. Bacteriol. 173 : 2576 - 2580 . (10.1128/jb.173.8.2576-2580.1991) / J. Bacteriol. / Three-dimensional structure of the nickelcontaining hydrogenase from Thiocapsa roseopersicina by Shermann M. B. (1991)
  43. Specka , U. , A. Spreinat , G. Antranikian , and F. Mayer . 1991 . Immunocytochemical identification and localization of active and inactive a-amylase and pullulanase in cells of Clostridium thermosulfurogenes EM1 . Appl. Environ. Microbiol. 57 : 1062 - 1069 . (10.1128/aem.57.4.1062-1069.1991) / Appl. Environ. Microbiol. / Immunocytochemical identification and localization of active and inactive a-amylase and pullulanase in cells of Clostridium thermosulfurogenes EM1 by Specka U. (1991)
  44. Sprott G. D. 1990. The F420-reducing hydrogenase of Methanospirillum hungatei strain GP1 p. 75-85. In J. B. B61aich M. Bruschi and J. L. Garcia (ed.) Microbiology and biochemistry of strict anaerobes involved in interspecies transfer. Plenum Press New York. (10.1007/978-1-4613-0613-9_6)
  45. Steigerwald , V. J. , T. D. Pihl , and J. N. Reeve . 1992 . Identification and isolation of the polyferredoxin from Methanobacterium thermoautotrophicum strain AH . Proc. Natl. Acad. Sci. USA 89 : 6929 - 6933 . (10.1073/pnas.89.15.6929) / Proc. Natl. Acad. Sci. USA / Identification and isolation of the polyferredoxin from Methanobacterium thermoautotrophicum strain AH by Steigerwald V. J. (1992)
  46. Towbin , H. , T. Staehelin , and J. Gordon . 1979 . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications . Proc. Natl. Acad. Sci. USA 76 : 4350 - 4354 . (10.1073/pnas.76.9.4350) / Proc. Natl. Acad. Sci. USA / Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications by Towbin H. (1979)
  47. Valentine , R. C. , B. M. Shapiro , and E. R. Stadtman . 1968 . Regulation of glutamine synthetase. XII. Electron microscopy of the enzyme from Escherichia coli . Biochemistry 7 : 2143 - 2152 . (10.1021/bi00846a017) / Biochemistry / Regulation of glutamine synthetase. XII. Electron microscopy of the enzyme from Escherichia coli by Valentine R. C. (1968)
  48. Wackett , L. P. , E. A. Hartwieg , J. A. King , W. H. Orme-Johnson , and C. T. Walsh . 1987 . Electron microscopy of nickel-containing methanogenic enzymes: methyl reductase and F420-reducing hydrogenase . J. Bacteriol. 169 : 718 - 727 . (10.1128/jb.169.2.718-727.1987) / J. Bacteriol. / Electron microscopy of nickel-containing methanogenic enzymes: methyl reductase and F420-reducing hydrogenase by Wackett L. P. (1987)
  49. Woo , G. , A. Wasserfallen , and R. S. Wolfe . 1993 . Methylviologen hydrogenase II, a new member of the hydrogenase family from Methanobacterium thermoautotrophicum AH . J. Bacteriol. 175 : 5970 - 5977 . (10.1128/jb.175.18.5970-5977.1993) / J. Bacteriol. / Methylviologen hydrogenase II, a new member of the hydrogenase family from Methanobacterium thermoautotrophicum AH by Woo G. (1993)
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Created 8 years, 9 months ago (Nov. 7, 2016, 4:37 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:20 p.m.)
Indexed 1 year, 10 months ago (Oct. 26, 2023, 7:18 p.m.)
Issued 30 years, 8 months ago (Dec. 1, 1994)
Published 30 years, 8 months ago (Dec. 1, 1994)
Published Print 30 years, 8 months ago (Dec. 1, 1994)
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@article{Braks_1994, title={Structural aspects and immunolocalization of the F420-reducing and non-F420-reducing hydrogenases from Methanobacterium thermoautotrophicum Marburg}, volume={176}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.176.24.7677-7687.1994}, DOI={10.1128/jb.176.24.7677-7687.1994}, number={24}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Braks, I J and Hoppert, M and Roge, S and Mayer, F}, year={1994}, month=dec, pages={7677–7687} }