Abstract
The localization of APS reductase and bisulfite reductase in Desulfovibrio gigas, D. vulgaris Hildenborough and D. thermophilus was studied by immunoelectron microscopy. Polyclonal antibodies were raised against the purified enzymes from each strain. Cells fixed with formaldehyde/glutaraldehyde were embedded and ultrathin sections were incubated with antibodies and subsequently labeled with protein A-gold. The bisulfite reductase in all three strains and APS reductase in d. gigas and D. vulgaris were found in the cytoplasm. The labeling of d. thermophilus with APS reductase antibodies resulted in a distribution of gold particles over the cytoplasmic membrane region. The localization of the two enzymes is discussed with respect to the mechanism and energetics of dissimilatory sulfate reduction.
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References
Akagi JM (1981) Dissimilatory sulfate reduction: mechanistic aspects. In: Bothe H, Trebst A (eds) Biology of inorganic nitrogen and sulfur. Springer, Berlin Heidelberg New York, pp 169–177
Aldrich HC, Beinborn DB, Bokranz M, Schönheit P (1987) Immunocytochemical localization of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum. Arch Microbiol 147:190–194
Badziong, W, Thauer RK (1980) Vectorial electron transport in Desulfovibrio vulgaris (Marburg) growing on hydrogen plus sulfate as sole energy source. Arch Microbiol 125:167–174
Barton LL, LeGall J, Peck HD (1972) Oxidative phosphorylation in the obligate anaerobe, Desulfovibrio gigas. In: San Pietro A, Gest H (eds) Horizons of bioenergetics. Academic Press, New York, pp 33–51
Blake MS, Johnston KH, Russel-Jones GJ, Gotchlich EC (1984) A rapid, sensitive method for detection of alkaline phosphataseconjugated anti-antibody on western blots. Anal Biochem 136:175–179
Bramlett RN, Peck HD (1975) Some physical and kinetic properties of adenylyl sulfate reductase from Desulfovibrio vulgaris. J Biol Chem 250:2979–2986
Brewer JM, Ashworth RB (1969) Disc electrophoresis. J Chem Educ 46:41–45
Carlemalm E, Garavito M, Villiger W (1982) Resin development for electron microcopy and analysis of embedding at low temperature. J Microscopy 126:123–143
Chambers LA, Trudinger PA (1975) Are thiosulfate and trithionate intermediates in dissimilatory sulfate reduction? J Bacteriol 123:36–40
Cohen J, Adautte A, Grandchamp S, Houdebine LM, Beisson J (1982) Immunocytochemical study of microtubular structures throughout the cell cycle of Paramecium. Biol Cell 44:35–44
Fauque GD, Czechowski MH, Kang L, Dervartanian DV, LeGall J (1986a) Purification of desulfofuscidin from a thermophilic strain of sulfate-reducing bacterium Desulfovibrio thermophilus. Abstract Ann Meet Am Soc Microbiol, Washington DC, K50 p 201
Fauque G, Czechowski MH, Kang-Lissolo L, DerVartanian DV, Moura JJG, Moura I, Lampreia J, Xavier AV, LeGall J (1986 b) Purification of adenylyl sulfate (APS) reductase and desulfofuscidin from a thermophilic sulfate reducer: Desulfovibrio thermophilus. Abstract Ann Meet Soc Ind Microbiol, San Francisco, p 92
Fauque GD, Moura I, Moura JJG, Xavier AV, Galliano N, LeGall J (1987) Isolation and characterization of a rubredoxin and a flavodoxin from Desulfovibrio desulfuricans Berre-Eau. FEBS Lett 215:63–67
Frens G (1973) Controlled nucleation for the regulation of particle size in monodisperse gold suspensions. Nature Phys Sci 241:20–22
Hansen TA (1988) Physiology of sulphate-reducing bacteria. Microbiol Sciences 5:81–84
Kohring GW, Mayer H, Mayer F (1985) Immunoelectron microscopic localization of the restriction endonuclease Eco RI in Escherichia coli BS 5. Eur J Cell Biol 37:1–6
Kremer DR, Hansen TA (1987) Glycerol and dihydroxyacetone dissimilation in Desulfovibrio strains. Arch Microbiol 147: 249–256
Lampreia J, Moura I, Fauque G, Xavier AV, LeGall J, Peck HD Jr., Moura JJG (1987) Adenylyl sulfate (APS) reductase form Desulfovibrio gigas. Rec Travaux Chim Pays-Bas 106:234
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227:680–685
Lee JP, Peck HD (1971) Purification of the enzyme reducing bisulfite to trithionate from Desulfovibrio gigas and its identification as desulfoviridin. Biochem Biophys Res Commun 45:583–589
Lee JP, LeGall J, Peck HD (1973) Isolation of assimilatory- and dissimilatory-type sulfite reductases from Desulfovibrio vulgaris. J Bacteriol 115:529–542
LeGall J, Fauque G (1988) Dissimilatory reduction of sulfur compounds. In: Zehnder AJB (ed) Biology of anaerobic microorganisms, Chapter 11. John Wiley and Sons, New York London (in press)
Lupton FS, Conrad R, Zeikus JG (1984) Physiological function of hydrogen metabolism during growth of sulfidogenic bacteria on organic substrates. J Bacteriol 159:843–849
Moor H (1964) Die Gefrier-Fixation lebender Zellen und ihre Anwendung in der Elektronen-Mikroskopie. Z Zellforsch 62:546–580
Moura JJG, Xavier AV, Hatchikian EC and LeGall J (1978) Structural control of the redox potentials and of the physiological activity by oligomerization of ferredoxin. FEBS Lett 89:177–179
Odom JM, Peck HD (1981a) Localization of dehydrogenases, reductases and electron transfer components in the sulfate-reducing bacterium Desulfovibrio gigas. J Bacteriol 147:161–169
Odom JM, Peck HD (1981b) Hydrogen cycling as a general mechanism for energy coupling in the sulfate reducing bacteria, Desulfovibrio sp. FEMS Microbiol Lett 12:47–50
Ossmer R, Mund T, Hartzell PL, Konheiser U, Kohring GW, Klein A, Wolfe RS, Gottschalk G, Mayer F (1986) Immunocytochemical localization of component C of the methylreductase system in Methanococcus voltae and Methanobacterium thermoautotrophicum. Proc Natl Acad Sci USA 83:5789–5792
Peck HD Jr., LeGall J (1982) Biochemistry of dissimilatory sulfate reduction. Phil Trans R Soc Lond B 298:443–446
Rohde M, Mayer F, Jacobitz S, Meyer O (1985) Attachment of CO dehydrogenase to the cytoplasmic membrane is limiting the respiratory rate of Pseudomonas carboxydovorans. FEMS Microbiol Lett 28:141–144
Roth J, Bendayan M, Orci L (1978) Ultrastructural localization of intracellular antigens by the use of protein A/gold complex. J Histochem Cytochem 26:1074–1081
Slot JW, Geuze HJ (1984) Gold markers for single and double immunolabelling of ultrathin cryosections. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elseviers Sci Publ, Amsterdam, pp 129–142
Stams AJM, Hansen TA (1982) Oxygen-labile L(+)lactate dehydrogenase activity in Desulfovibrio HL21. FEMS Microbiol Lett 13:389–394
Stams AJM, Veenhuis M, Weenk GH, Hansen TA (1983) Occurrence of polyglucose as a storage polymer in Desulfovibrio species and Desulfobulbus propionicus. Arch Microbiol 136: 54–59
Starkey RL (1938) A study on spore formation and other morphological characteristics of Vibrio desulfuricans. Arch Mikrobiol 8:268–304
Stille W, Trüper HG (1984) Adenylylsulfate reductase in some new sulfate-reducing bacteria. Arch Microbiol 137:145–150
Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180
Trüper HG, Fischer U (1982) Anaerobic oxidation of sulphur compounds as electron donors for bacterial photosynthesis. Phil Trans R Soc Lond B 298:529–542
Turner BM (1986) Use of alkaline phosphatase-conjugated antibodies for detection of protein antigens in nitrocellulose filters. In: Langone JJ, Vunakis HV (eds) Methods in enzymology 141:848–855
Widdel F (1988) Microbiology and ecology of sulfate and sulfurreducing bacteria. In: Zehnder AJB (ed) Biology of anaerobic microorganisms, chapter 10. John Wiley and Sons, New York London (in press)
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Kremer, D.R., Veenhuis, M., Fauque, G. et al. Immunocytochemical localization of APS reductase and bisulfite reductase in three Desulfovibrio species. Arch. Microbiol. 150, 296–301 (1988). https://doi.org/10.1007/BF00407795
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DOI: https://doi.org/10.1007/BF00407795