Abstract
The indirect immunoferritin labeling method was used to localize the membrane-bound respiratory nitrate reductase in membrane vesicles and protoplasts or spheroplasts of Bacillus licheniformis and Klebsiella aerogenes, respectively. For a comparison of the labeling of the various vesicle preparations, which differed not only in size but also in the percentage of inside-out orientation, a quantification of the results was needed to circumvent the problem of non-specifically bound ferritin. From the results the sidedness of the nitrate reductase in the cytoplasmic membrane of the abovementioned bacteria was determined as being cytoplasmic in B. licheniformis and as transmembranous in K. aerogenes.
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Abbreviations
- PBS:
-
phosphate buffered saline
- IgG:
-
immunoglobulin G
References
Carraway, K. L.: Covalent labeling of membranes. Biochim. Biophys. Acta 415, 379–410 (1975)
Davis, W. C.: Use of antibodies for localization of components on membranes, in Methods in Enzymology (S. Fleischer, L. Packer, eds.), Vol. 32, pp. 60–70. New York: Academic Press 1974
Fowler, S., Remacle, J., Trouet, A., Beaufay, H., Berthet, J., Wibo, M., Hauser, P.: Analytical study of microsomes and isolated subcellular membranes from rat liver. J. Cell Biol. 71, 535–550 (1976)
Ghosh, A., Ghosh, B. K.: Immuno-electron microscopic localization of penicillinase in Bacillus licheniformis. J. Bacteriol. 137, 1374–1385 (1979)
Kalb, V. F., Jr., Bernlohr, R. W.: A new spectrophotometric assay for protein in cell extracts. Anal. Biochem. 82, 362–371 (1977)
Kawamura, A., Jr.: Fluorescent antibody techniques and their applications. Baltimore, M.D.: University Park Press 1969
Kishida, Y., Olsen, B. R., Berg, R. A., Prockop, D. J.: Two improved methods for preparing ferritin-antibody conjugates for electronmicroscopy. J. Cell Biol. 64, 331–339 (1975)
Macalister, T. J., Irvin, R. T., Costerton, J. W.: Immunocytological investigation of protein synthesis in Escherichia coli. J. Bacteriol. 130, 329–338 (1977)
Matsuura, S., Fujii-Kuriyama, Y., Tashiro, Y.: Immunoelectronmicroscope localization of cytochrome P-450 on microsomes and other membrane structures of rat hepatocytes. J. Cell Biol. 78, 503–519 (1978)
Matsuura, S., Fujii-Kuriyama, Y., Tashiro, Y.: Quantitative immunoelectron microscopic analyses of the distribution of cytochrome P-459 molecules on rat liver microsomes. J. Cell Sci. 36, 413–435 (1979)
Morimoto, T., Matsuura, S., Sasaki, S., Tashiro, Y., Omura, T.: Immunochemical and immuno-electronmicroscopic studies on localization of NADH-cytochrome c reductase on rat liver microsomes. J. Cell Biol. 68, 189–210 (1976)
Nicolson, G. L., Painter, R. G.: Anionic sites of human erythrocyte membranes. J. Cell Biol. 59, 395–406 (1973)
Oppenheim, J. D., Salton, M. R. J.: Localization and distribution of Micrococcus leisodeikticus membrane ATPase determined by ferritin labeling. Biochim. Biophys. Acta 298, 297–322 (1973)
Otto, H., Takamiya, H., Vogt, A.: A two-stage method for cross linking antibody globulin to ferritin by glutaraldehyde.J. Immunol. Meth. 3, 137–146 (1973)
The, T. H., Feltkamp, T. E. W.: Conjugations of FITC to antibodies. Immunology 18, 875–881 (1970)
Van't Riet, J., Planta, R. J.: Purification, structure and properties of the respiratory nitrate reductase of Klebsiella aerogenes. Biochim. Biophys. Acta 379, 81–94 (1975)
Van't Riet, J., Wientjes, F. B., van Doorn, J., Planta, R. J.: Purification and properties of the respiratory nitrate reductase of Bacillus licheniformis. Biochim. Biophys. Acta 576, 347–360 (1979)
Wientjes, F. B., Kolk, A. H. J., Nanninga, N., Van't Riet, J.: Respiratory nitrate reductase: its localization in the cytoplasmic membrane of Klebsiella aerogenes and Bacillus licheniformis. Eur. J. Biochem. 95, 61–67 (1979a)
Wientjes, F. B., Van't Riet, J., Nanninga, N.: Formation of insideout vesicles of Bacillus licheniformis: dependence on buffer composition and lysis procedure. Biochim. Biophys. Acta 553, 213–223 (1979b)
Wientjes, F. B.: Localization of respiratory nitrate reductase in the cytoplasmic membrane of Bacillus licheniformis and Klebsiella aerogenes. Ph.D. Thesis Free University, Amsterdam (1979)
Witholt, B., Boekhout, M., Brock, M., Kingma, J., Van Heerikhuizen, H., De Leij, L.: An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal. Biochem. 74, 160–170 (1976)
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Wientjes, F.B., Riet, J.v. & Nanninga, N. Immunoferritin labeling of respiratory nitrate reductase in membrane vesicles of Bacillus licheniformis and Klebsiella aerogenes . Arch. Microbiol. 127, 39–46 (1980). https://doi.org/10.1007/BF00414353
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DOI: https://doi.org/10.1007/BF00414353