Skip to main content

Advertisement

SpringerLink
Log in

Mixed-function oxidases of 25-hydroxycholecalciferol in isolated chick kidney glomeruli: Evidence for nuclear localization

  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Summary

Pure chick kidney glomeruli and proximal tubular fragments have been isolated by graded sieving through nylon screens. Electron micrographs revealed that, in distinct contrast to proximal epithelial cells, the glomerular epithelial and endothelial cells are essentially devoid of mitochondria. Glomeruli as well as proximal tubular fragments contain the 1α- and 24R-hydroxylases of 25-hydroxycholecalciferol. The level of 1α-hydroxylase activity was the same in both segments of the nephron. However, the tubular fragments contained twice the 24R-hydroxylase activity found in glomeruli. Glomerular nuclei were purified by sucrose gradient sedimentation and used to confirm the association of the 1α-hydroxylase with this kidney organelle. Almost all of the glomerular 1α-hydroxylase activity was found in the nuclear fraction. Two metabolites, which are produced predominantly by the nuclei, are designated N-1 and N-2. Their structural identity remains unknown. The novel presence of the 1α-hydroxylase in the glomerulus may be important in defining the etiology of bone diseases in patients with glomerulonephritis and similar disorders.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ponchon, G., Kennan, A.L., DeLuca, H.F.: Activation of vitamin D by the liver, J. Clin. Invest.48:2032–2037, 1969

    Article  CAS  PubMed  Google Scholar 

  2. Fraser, D.R., Kodicek, E.: Unique biosynthesis by kidney of a biologically active vitamin D metabolite, Nature228:764–766, 1970

    Article  CAS  PubMed  Google Scholar 

  3. Gray, R.W., Omdahl, J.L., Ghazarian, J.G., DeLuca, H.F.: 25-Hydroxycholecalciferol-1-hydroxylase. Subcellular location and properties, J. Biol. Chem.247:7528–7532, 1972

    CAS  PubMed  Google Scholar 

  4. Ghazarian, J.G.: The 1α- and 24-hydroxylases of 25-hydroxycholecalciferol: One or two enzymes? In A.W. Norman, K. Schaefer, D.V. Herrath, H. G. Grigoleit, J. W. Coburn, H. F. DeLuca, E. B. Mawer, T. Suda (eds): Vitamin D Basic Research and its Clinical Application, pp. 587–596. Walter de Gruyter, Berlin, 1979

    Google Scholar 

  5. Kulkoski, J.A., Chan, T., Martinez, J., Ghazarian, J.G.: Modulation of 25-hydroxyvitamin D3-24-hydroxylase by aminophylline: A cytochrome P-450 monooxygenase system, Biochem. Biophys. Res. Common.90:50–57, 1979

    Article  Google Scholar 

  6. Ghazarian, J.G., Garancis, J.C.: Immunofluorescent localization of 25-hydroxyvitamin-D3-1α-hydroxylase ferredoxin in the renal tissue of chick, J. Histochem. Cytochem.27:1041–1045, 1979

    CAS  PubMed  Google Scholar 

  7. Kulkoski, J.A., Peterson, B.L., Elcombe, B., Winkelhake, J.L., Ghazarian, J.G.: Ferredoxin of 25-hydroxyvitamin D3-1α-hydroxylase: Anatomical distribution in the chick as determined by double-antibody radioimmunoassay, FEBS Lett.99:183–188, 1979

    Article  CAS  PubMed  Google Scholar 

  8. Kulkoski, J.A., Ghazarian, J.G.: Purification and characterization of the ferredoxin component of 25-hydroxycholecalciferol 1α-hydroxylase, Biochem. J.177:673–678, 1979

    CAS  PubMed  Google Scholar 

  9. Brunette, M.G., Chan, M., Ferriere, C. Roberts, K.D.: Site of 1,25(OH)2D3 synthesis in the kidney, Nature276:287–289, 1978

    Article  CAS  PubMed  Google Scholar 

  10. Dencker, L., Tjalve, H.: Studies on the renal uptake of vitamin D3 in the mouse and the quail, Acta Physiol. Scand.95:407–416, 1975

    Article  CAS  PubMed  Google Scholar 

  11. Ghazarian, J.G., Weber, J.L., Sealy, R.C.: Solubilization of chick kidney microsomal cytochrome P-450: Electron paramagnetic resonance studies of ligand interactions, FEBS Lett.86:213–218, 1978

    Article  CAS  PubMed  Google Scholar 

  12. Norgaard, J.O.R.: A new method for the isolation of ultra-structually preserved glomeruli, Kidney International9:278–285, 1976

    CAS  PubMed  Google Scholar 

  13. Lawson, D.E.M., Wilson, P.W., Barker, D.C., Kodicek, E.: Isolation of chick intestinal nuclei—Effect of vitamin D3 on nuclear metabolism, Biochem. J.115:263–268, 1969

    CAS  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.193:265–275, 1951

    CAS  PubMed  Google Scholar 

  15. Ghazarian, J.G., DeLuca, H.F.: 25-Hydroxycholecalciferol-1-hydroxylase: A specific requirement for NADPH and a hemoprotein component in chick kidney mitochondria, Arch. Biochem. Biophys.160:63–72, 1974

    CAS  PubMed  Google Scholar 

  16. Bligh, E.G., Dyer, M.J.: A rapid method of total lipid extraction and purification, Can. J. Biochem. Physiol.37:911–917, 1959

    CAS  PubMed  Google Scholar 

  17. Fong, J.S.C., Drummond, K.N.: Method for preparation of glomeruli for metabolic studies, J. Lab. Clin Med.71:1034–1039, 1968

    CAS  PubMed  Google Scholar 

  18. Gang, N.F.: A rapid method for the isolation of glomeruli from the human kidney, Am. J. Clin. Pathol.53:267–269, 1970

    CAS  PubMed  Google Scholar 

  19. Misra, R.P.: Isolation of glomeruli from mammalian kidneys by graded sieving, Am. J. Clin. Pathol.58:135–139, 1972

    CAS  PubMed  Google Scholar 

  20. Boyle, I.T., Gray, R.W., DeLuca, H.F.: Regulation by calcium ofin vivo synthesis of 1,25-Dihydroxycholecalciferol and 21,25-Dihydroxycholecalciferol, Proc. Nat. Acad. Sci.68:2131–2134, 1971

    CAS  PubMed  Google Scholar 

  21. Tanaka, Y., DeLuca, H.F., Kobayashi, Y., Taguchi, T., Ikekawa, N., Morisaki, M.: Biological activity of 24,24-difluoro-25-hydroxyvitamin D3—Effect of blocking of 24-hydroxylation on the functions of vitamin D, J. Biol. Chem.254:7163–7167, 1979

    CAS  PubMed  Google Scholar 

  22. Gray, R.W., Weber, H.P., Dominguez, J.H., Lemann, J. Jr.: The metabolism of vitamin D3 and 25-hydroxyvitamin D3 in normal and anephoic humans, J. Clin. Endo. Metab.39:1045–1056, 1974

    Article  CAS  Google Scholar 

  23. Avioli, L.V., Lee, S.W., McDonald, J.E., Lund, J., DeLuca, H.F.: Metabolism of vitamin D3-3H in human subjects: Distinction in blood, bile, feces, and urine. J. Clin. Invest.46:983–992, 1967

    CAS  PubMed  Google Scholar 

  24. Fahl, W.E., Jefcoate, C.R., Kasper, C.B.: Characteristics of benzo(a)pyrene metabolism and cytochrome P-450 heterogeneity in rat liver nuclear envelope and comparison to microsomal membrane, J. Biol. Chem.253:3106–3113, 1978

    CAS  PubMed  Google Scholar 

  25. Kasper, C.B.: Biochemical distinctions between the nuclear and microsomal membranes from rat hepatocytes—The effect of phenobarbital administration, J. Biol. Chem.246:577–581, 1971

    CAS  PubMed  Google Scholar 

  26. Khandwala, A.S., Kasper, C.B.: Preferential induction of aryl hydroxylase activity in rat liver nuclear envelope by 3-methylcholanthrene, Biochem. Biophys. Res. Commun.54:1241–1246, 1973

    Article  CAS  PubMed  Google Scholar 

  27. Rogan, E.G., Cavalieri, E.: 3-Methylcholanthrene-inducible binding of aromatic hydrocarbons to DNA in purified rat liver nuclei, Biochem. Biophys. Res. Commun.58:1119–1126, 1974

    Article  CAS  PubMed  Google Scholar 

  28. Rogan, E.G., Cavalieri, E.: Differences between nuclear and microsomal cytochrome P-450 in uninduced and induced rat liver, Molec. Pharmac.14:215–219, 1978

    CAS  Google Scholar 

  29. Rogan, E.G., Mailander, P., Cavalieri, E.: Metabolic activation of aromatic hydrocarbons in purified rat liver nuclei: Induction of enzyme activities and binding to DNA with and without monooxygenase-catalyzed formation of active oxygen, Proc. Natl. Acad. Sci.73:457–461, 1976

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, Medical College of Wisconsin, 53226, Milwaukee, Wisconsin, USA

    Eric W. Peterson & Jacob G. Ghazarian

  2. Department of Pathology, Medical College of Wisconsin, 53226, Milwaukee, Wisconsin, USA

    John C. Garancis

Authors
  1. Eric W. Peterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jacob G. Ghazarian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John C. Garancis
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Peterson, E.W., Ghazarian, J.G. & Garancis, J.C. Mixed-function oxidases of 25-hydroxycholecalciferol in isolated chick kidney glomeruli: Evidence for nuclear localization. Calcif Tissue Int 33, 19–25 (1981). https://doi.org/10.1007/BF02409408

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02409408

Key words

Search

Navigation