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Histochemistry

, Volume 95, Issue 1, pp 37–42 | Cite as

Antigenic and catalytic disparity in the distribution of cytochrome P-450-dependent 25-hydroxyvitamin D3-1α- and 24-hydroxylases

  • K. Takezawa
  • B. Moorthy
  • M. L. Mandel
  • J. C. Garancis
  • J. G. Ghazarian
Article

Summary

Chick 25-hydroxyvitamin D3-1α-hydroxylase, a cytochrome P-450 monooxygenase with a molecular weight of 57 kDa, can be isolated as described by Mandel et al. (1990b). Under normal physiological circumstances, it occurs exclusively in kidney mitochondria. An isozyme of the 1α-hydroxylase, known as the 24-hydroxylase, which uses the same substrate to yield an isomeric product, is also a cytochrome P-450 monooxygenase, has a molecular weight of 55 kDa, and likewise occurs in kidney mitochondria. The amino-terminal sequences of the first 10 residues of the two isozymes are 100% homologous. Monoclonal antibodies of the IgM class raised against the 1α-hydroxylase, which quantitatively discriminate against other P-450 cytochromes of mitochondrial or microsomal origin, recognize and interact with the 24-hydroxylase as an antigen. In the present study we show that the intestine, which is the only non-renal tissue with demonstrable 24-hydroxylase activity, gives a positive peroxidase-antiperoxidase immunohistochemical reaction using the monoclonal antibodies against the 1α-hydroxylase. The reactions revealed that the antigen in the kidney is restricted to the cortical proximal tubular cells while in the intestine, the antigen is localized in the enterocytes of the villi. In kidney medullary or intestinal crypt cells, or in liver, heart and lung tissues where 1α-hydroxylase or 24-hydroxylase activity could not be detected using cell or tissue homogenates, the immunohistochemical reactions were also negative. Since it has been reported that chick embryonic intestine possesses 1α-hydroxylase activity that is absent in the mature intestine, our results would suggest that the mature intestinal 24-hydroxylase represents a modified 1α-hydroxylase as a consequence of developmentally imposed requirements regulating calcium homeostatic activity in this tissue. The difference in the molecular weights of the two enzymes would indicate either genomic processing prior to the translation of their respective mRNAs, or a post-translational processing of the larger 1α-hydroxylase to the smaller 24-hydroxylase.

Keywords

Crypt Cell Proximal Tubular Cell Immunohistochemical Reaction Intestinal Crypt Kidney Mitochondrion 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • K. Takezawa
    • 1
    • 3
  • B. Moorthy
    • 1
  • M. L. Mandel
    • 1
  • J. C. Garancis
    • 2
  • J. G. Ghazarian
    • 1
  1. 1.Department of BiochemistryMedical College of WisconsinMilwaukeeeUSA
  2. 2.Department of PathologyMedical College of WisconsinMilwaukeeeUSA
  3. 3.Department of Internal MedicineToho University School of MedicineTokyoJapan

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