Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Aug;159(2):668–677. doi: 10.1128/jb.159.2.668-677.1984

Subcellular localization of alkaline phosphatase in Bacillus licheniformis 749/C by immunoelectron microscopy with colloidal gold.

G Tinglu, A Ghosh, B K Ghosh
PMCID: PMC215696  PMID: 6746574

Abstract

Subcellular distribution of the alkaline phosphatase of Bacillus licheniformis 749/C was determined by an immunoelectron microscopy method. Anti-alkaline phosphatase antibody labeled with 15- to 18-nm colloidal gold particles (gold-immunoglobulin G [IgG] complex) were used for the study. Both the plasma membrane and cytoplasmic material were labeled with the gold-IgG particles. These particles formed clusters in association with the plasma membrane; in contrast, in the cytoplasm the particles were largely dispersed, and only a few clusters were found. The gold-IgG binding was quantitatively estimated by stereological analysis of labeled, frozen thin sections. This estimation of a variety of control samples showed that the labeling was specific for the alkaline phosphatase. Cluster formation of the gold-IgG particles in association with the plasma membrane suggests that existence of specific alkaline phosphatase binding sites (receptors) in the plasma membrane of B. licheniformis 749/C.

Full text

PDF
668

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bayer M. E. Structural and functional evidence of cooperativity between membranes and cell wall in bacteria. Int Rev Cytol Suppl. 1981;12:39–70. doi: 10.1016/b978-0-12-364373-5.50012-3. [DOI] [PubMed] [Google Scholar]
  2. Bendayan M., Zollinger M. Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem. 1983 Jan;31(1):101–109. doi: 10.1177/31.1.6187796. [DOI] [PubMed] [Google Scholar]
  3. Christensen A. K. Frozen thin sections of fresh tissue for electron microscopy, with a description of pancreas and liver. J Cell Biol. 1971 Dec;51(3):772–804. doi: 10.1083/jcb.51.3.772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ghosh A., Ghosh B. K. Changes in the membrane bound alkaline phosphatase of glucose and lactate grown vegetative cells of Bacillus subtilis SB15. Biochem Biophys Res Commun. 1972 Nov 15;49(4):906–915. doi: 10.1016/0006-291x(72)90298-7. [DOI] [PubMed] [Google Scholar]
  5. Ghosh A., Ghosh B. K. Immunoelectron microscopic localization of penicillinase in Bacillus licheniformis. J Bacteriol. 1979 Mar;137(3):1374–1385. doi: 10.1128/jb.137.3.1374-1385.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ghosh A., Vallespir S., Ghosh B. K. Specificity of subcellular distribution of alkaline phosphatase in Bacillus licheniformis 749/C. Can J Microbiol. 1984 Jan;30(1):113–125. doi: 10.1139/m84-019. [DOI] [PubMed] [Google Scholar]
  7. Ghosh B. K., Nanninga N. Polymorphism of the mesosome in Bacillus licheniformis (749/C and 749). Influence of chemical fixation monitored by freeze-etching. J Ultrastruct Res. 1976 Jul;56(1):107–120. doi: 10.1016/s0022-5320(76)80144-x. [DOI] [PubMed] [Google Scholar]
  8. Ghosh B. K., Wouters J. T., Lampen J. O. Distribution of the sites of alkaline phosphatase(s) activity in vegetative cells of Bacillus subtilis. J Bacteriol. 1971 Nov;108(2):928–937. doi: 10.1128/jb.108.2.928-937.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ghosh R., Ghosh A., Ghosh B. K. Properties of the membrane-bound alkaline phosphatase from glucose- and lactate-grown cells of Bacillus subtilis SB 15. J Biol Chem. 1977 Oct 10;252(19):6813–6822. [PubMed] [Google Scholar]
  10. Handley D. A., Arbeeny C. M., Witte L. D., Chien S. Colloidal gold--low density lipoprotein conjugates as membrane receptor probes. Proc Natl Acad Sci U S A. 1981 Jan;78(1):368–371. doi: 10.1073/pnas.78.1.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horisberger M., Rosset J., Bauer H. Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope. Experientia. 1975 Oct 15;31(10):1147–1149. doi: 10.1007/BF02326761. [DOI] [PubMed] [Google Scholar]
  12. Horisberger M., Vonlanthen M. Multiple marking of cell surface receptors by gold granules: simultaneous localization of three lectin receptors on human erythrocytes. J Microsc. 1979 Jan;115(1):97–102. doi: 10.1111/j.1365-2818.1979.tb00155.x. [DOI] [PubMed] [Google Scholar]
  13. Kumar R., Ghosh A., Ghosh B. K. Alkaline phosphatase secretion-negative mutant of Bacillus licheniformis 749/C. J Bacteriol. 1983 May;154(2):946–954. doi: 10.1128/jb.154.2.946-954.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Michaelis S., Beckwith J. Mechanism of incorporation of cell envelope proteins in Escherichia coli. Annu Rev Microbiol. 1982;36:435–465. doi: 10.1146/annurev.mi.36.100182.002251. [DOI] [PubMed] [Google Scholar]
  16. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Roth J., Bendayan M., Orci L. Protein A-gold complex for postembedding staining of intracellular antigens. Acta Histochem Suppl. 1980;22:269–273. [PubMed] [Google Scholar]
  18. Roth J., Binder M. Coloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques. J Histochem Cytochem. 1978 Mar;26(3):163–169. doi: 10.1177/26.3.632554. [DOI] [PubMed] [Google Scholar]
  19. Tokuyasu K. T., Singer S. J. Improved procedures for immunoferritin labeling of ultrathin frozen sections. J Cell Biol. 1976 Dec;71(3):894–906. doi: 10.1083/jcb.71.3.894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Walter P., Ibrahimi I., Blobel G. Translocation of proteins across the endoplasmic reticulum. I. Signal recognition protein (SRP) binds to in-vitro-assembled polysomes synthesizing secretory protein. J Cell Biol. 1981 Nov;91(2 Pt 1):545–550. doi: 10.1083/jcb.91.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weibel E. R. Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol. 1969;26:235–302. doi: 10.1016/s0074-7696(08)61637-x. [DOI] [PubMed] [Google Scholar]
  22. de Bruijn W. C., Emeis J. J., Vermeer B. J. The application of analytical electron microscopy in the localization of individual LDL-binding sites on cell surfaces. Artery. 1980;8(3):281–287. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES