1887

Abstract

pellicle, separated by sucrose density gradient centrifugation, had a high cobalamin (Cb1) binding activity. Of the Cb1 binding protein 65% was solubilized from pellicle by using 0·1% SDS/2 -urea and the residual 35% by using 1% (w/v) SDS. Both of the Cb1 binding protein fractions showed a broad pH dependence for the binding of Cb1. No evidence for the involvement of SH-groups or metal ions in Cb1 binding was obtained. The values for cyanocobalamin of the proteins solubilized with 0·1% SDS/2 -urea and with 1% SDS were 0·22 and 0·19 n, respectively. The of a Cb1 binding polypeptide of each protein was estimated to be 38000 by SDS-PAGE. The Cb1 binding protein solubilized with 0·1% SDS/2 -urea was purified to homogeneity. Inhibition experiments on Cb1 uptake in cells, using an antibody against the Cb1 binding protein solubilized with 0·1% SDS/2 -urea, showed that the Cb1 binding proteins solubilized with 0·1% SDS/2 -urea and 1% SDS take part in the slower secondary phase and in the initial rapid phase of Cb1 uptake in , respectively.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-1-67
1988-01-01
2021-10-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/1/mic-134-1-67.html?itemId=/content/journal/micro/10.1099/00221287-134-1-67&mimeType=html&fmt=ahah

References

  1. Allen R.H., Majerus P.W. 1972; Isolation of vitamin B12-binding proteins using affinity chromatography III. Purification and properties of human plasma transcobalamin II. Journal of Biological Chemistry 247:7709–7717
    [Google Scholar]
  2. Allen R.H., Mehlman C.S. 1973; Isolation of gastric vitamin B12-binding proteins using affinity chromatography I. Purification and properties of human intrinsic factor. Journal of Biological Chemistry 248:3660–3669
    [Google Scholar]
  3. Bradbeer C. 1971; Transport of vitamin B12 in Ochromonas malhamensis. Archives of Biochemistry and Biophysics 144:184–192
    [Google Scholar]
  4. Bradbeer C., Woodrow M.L. 1976; Transport of vitamin B12 in Escherichia coli: energy dependence. Journal of Bacteriology 128:99–104
    [Google Scholar]
  5. Bradbeer C., Kenley J.S., Dimasi D.R., Leighton M. 1978; Transport of vitamin B12 in Escherichia coli Corrinoid specificities of the periplasmic B12-binding protein and of energy-dependent B12 transport. Journal of Biological Chemistry 253:1347–1352
    [Google Scholar]
  6. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  7. Cotter R., Rothenberg S.P., Weiss J.P. 1977; Purification of the intestinal receptor for intrinsic factor by affinity chromatography. Biochimica et biophysica acta 490:19–26
    [Google Scholar]
  8. Davis B.D. 1964; Disc electrophoresis II. Method and application to human protein. Annals of the NewYork Academy of Sciences 121:404–427
    [Google Scholar]
  9. Digirolama P.M., Bradbeer C. 1971; Transport of vitamin B12 in Escherichia coli. Journal of Bacteriology 106:745–750
    [Google Scholar]
  10. Hippe E., Haber E., Oleson H. 1971; Nature of vitamin B12 binding II. Steric orientation of vitamin B12 on binding and number of combining sites of human intrinsic factor and the transcobalamins. Biochimica et biophysica acta 243:75–82
    [Google Scholar]
  11. Hofmann C., Bouck S.H. 1976; Immunological and structural evidence of patterned intussusceptive surface growth in a unicellular organism. Journal of Cell Biology 69:693–715
    [Google Scholar]
  12. Isegawa Y., Nakano Y., Kitaoka S. 1985; Conversion and distribution of cobalamin in Euglena gracilis Z, with special reference to its location and probable function within chloroplasts. Plant Physiology 76:814–818
    [Google Scholar]
  13. Kadner R.J. 1978; Repression of synthesis of the vitamin B12 receptor in Escherichia coli. Journal of Bacteriology 136:1050–1057
    [Google Scholar]
  14. Kenley J.S., Leighton M., Bradbeer C. 1978; Transport of vitamin B12 in Escherichia coli Corrinoid specificity of the outer membrane receptor. Journal of Biological Chemistry 253:1341–1346
    [Google Scholar]
  15. Koren L.E., Hutner S.H. 1967; High-yield media for photosynthesizing Euglena gracilis. Journal of Protozoology 14: (Supplement) 17:
    [Google Scholar]
  16. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  17. Marcoullis G., Grasbeck R. 1977; Solubilized intrinsic factor receptor from pig ileum and its characteristics. Biochimica et biophysica acta 496:36–51
    [Google Scholar]
  18. Ross G.I.M. 1952; Vitamin B12 assay in body fluids using Euglena gracilis. Journal of Clinical Pathology 5:250–256
    [Google Scholar]
  19. Sabet S.T., Schnaitman C.A. 1973; Purification and properties of the colicin E 3 receptor of Escherichia coli. Journal of Biological Chemistry 248:1797–1806
    [Google Scholar]
  20. Sarhan F., Houde M., Cheneval J.P. 1980; The role of vitamin B12 binding in the uptake of the vitamin by Euglena gracilis. Journal of Protozoology 27:235–238
    [Google Scholar]
  21. Seligman P.A., Allen R.H. 1978; Characterization of the receptor for transcobalamin II isolated from human placenta. Journal of Biological Chemistry 253:1766–1772
    [Google Scholar]
  22. Taylor R.T., Norrell S.A., Hanna M.L. 1972; Uptake of cyanocobalamin by Escherichia coli B: Some characteristics and evidence for a binding protein. Archives of Biochemistry and Biophysics 148:366–381
    [Google Scholar]
  23. Youngdahl-Turner P., Mellman I.S., Allen R.H., Rosenberg L.E. 1979; Protein mediated vitamin uptake. Adsorptive endocytosis of the transcobalamin II-cobalamin complex by cultured human fibroblasts. Experimental Cell Research 118:127–134
    [Google Scholar]
  24. Varma T.N.S., Abraham A., Hansen I.A. 1961; Accumulation of 58Co vitamin B12 by Euglena gracilis. Journal of Protozoology 8:212–216
    [Google Scholar]
  25. White J.C., Digirolamo P.M., Fu M.L., Preston Y.A., Bradbeer C. 1973; Transport of vitamin B! 2 in Escherichia coli Location and properties of the initial B12-binding site. Journal of Biological Chemistry 248:3978–3986
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-1-67
Loading
/content/journal/micro/10.1099/00221287-134-1-67
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error