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. 1983 Jun 1;211(3):553–558. doi: 10.1042/bj2110553

Characterization of human foetal intestinal alkaline phosphatase. Comparison with the isoenzymes from the adult intestine and human tumour cell lines.

C M Behrens, C A Enns, H H Sussman
PMCID: PMC1154399  PMID: 6882358

Abstract

The molecular structure of human foetal intestinal alkaline phosphatase was defined by high-resolution two-dimensional polyacrylamide-gel electrophoresis and amino acid inhibition studies. Comparison was made with the adult form of intestinal alkaline phosphatase, as well as with alkaline phosphatases isolated from cultured foetal amnion cells (FL) and a human tumour cell line (KB). Two non-identical subunits were isolated from the foetal intestinal isoenzyme, one having same molecular weight and isoelectric point as placental alkaline phosphatase, and the other corresponding to a glycosylated subunit of the adult intestinal enzyme. The FL-cell and KB-cell alkaline phosphatases were also found to contain two subunits similar to those of the foetal intestinal isoenzyme. Characterization of neuraminidase digests of the non-placental subunit showed it to be indistinguishable from the subunits of the adult intestinal isoenzyme. This implies that no new phosphatase structural gene is involved in the transition from the expression of foetal to adult intestinal alkaline phosphatase, but that the molecular changes involve suppression of the placental subunit and loss of neuraminic acid from the non-placental subunit. Enzyme-inhibition studies demonstrated an intermediate response to the inhibitors tested for the foetal intestinal, FL-cell and KB-cell isoenzymes when compared with the placental, adult intestinal and liver forms. This result is consistent with the mixed-subunit structure observed for the former set of isoenzymes. In summary, this study has defined the molecular subunit structure of the foetal intestinal form of alkaline phosphatase and has demonstrated its expression in a human tumour cell line.

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Selected References

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

  1. Badger K. S., Sussman H. H. Structural evidence that human liver and placental alkaline phosphatase isoenzymes are coded by different genes. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2201–2205. doi: 10.1073/pnas.73.7.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benham F. J., Harris H. Human cell lines expressing intestinal alkaline phosphatase. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4016–4019. doi: 10.1073/pnas.76.8.4016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  4. Fishman W. H., Inglis N. I., Stolbach L. L., Krant M. J. A serum alkaline phosphatase isoenzyme of human neoplastic cell origin. Cancer Res. 1968 Jan;28(1):150–154. [PubMed] [Google Scholar]
  5. Fishman W. H., Sie H. G. Organ-specific inhibition of human alkaline phosphatase isoenzymes of liver, bone, intestine and placenta; L-phenylalanine, L-tryptophan and L homoarginine. Enzymologia. 1971 Sep 30;41(3):141–167. [PubMed] [Google Scholar]
  6. Gogolin K. J., Wray L. K., Slaughter C. A., Harris H. A monoclonal antibody that reacts with nonallelic enzyme glycoproteins. Science. 1982 Apr 2;216(4541):59–61. doi: 10.1126/science.6175022. [DOI] [PubMed] [Google Scholar]
  7. Gottlieb A. J., Sussman H. H. Human placental alkaline phosphatase: molecular weight and subunit structure. Biochim Biophys Acta. 1968 Jun 26;160(2):167–171. doi: 10.1016/0005-2795(68)90083-4. [DOI] [PubMed] [Google Scholar]
  8. Hamilton T. A., Tin A. W., Sussman H. H. Alkaline phosphatase isoenzyme expression in Chang liver cells. Exp Cell Res. 1979 Aug;122(1):31–38. doi: 10.1016/0014-4827(79)90557-3. [DOI] [PubMed] [Google Scholar]
  9. Hamilton T. A., Tin A. W., Sussman H. H. Regulation of alkaline phosphatase expression in human choriocarcinoma cell lines. Proc Natl Acad Sci U S A. 1979 Jan;76(1):323–327. doi: 10.1073/pnas.76.1.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harkness E. R. Studies on human placental alkaline phosphatase. I. Purification and crystallization. Arch Biochem Biophys. 1968 Aug;126(2):503–512. doi: 10.1016/0003-9861(68)90435-9. [DOI] [PubMed] [Google Scholar]
  11. Higashino K., Kudo S., Otani R., Yamamura Y. A hepatoma-associated alkaline phosphatase, the Kasahara isozyme, compared with one of the isozymes of FL amnion cells. Ann N Y Acad Sci. 1975 Aug 22;259:337–346. doi: 10.1111/j.1749-6632.1975.tb25430.x. [DOI] [PubMed] [Google Scholar]
  12. Ludueña M. A., Sussman H. H. Characterization of KB cell alkaline phosphatase. Evidence of similarity to placental alkaline phosphatase. J Biol Chem. 1976 May 10;251(9):2620–2628. [PubMed] [Google Scholar]
  13. McKenna M. J., Hamilton T. A., Sussman H. H. Comparison of human alkaline phosphatase isoenzymes. Structural evidence for three protein classes. Biochem J. 1979 Jul 1;181(1):67–73. doi: 10.1042/bj1810067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miki K., Oda T., Suzuki H., Niwa H., Endo Y., Iino S., Miyazaki J., Hirano K., Sugiura M. Human fetal organ alkaline phosphatases. Clin Chim Acta. 1978 Apr 17;85(2):115–124. doi: 10.1016/0009-8981(78)90230-9. [DOI] [PubMed] [Google Scholar]
  15. Mulivor R. A., Hannig V. L., Harris H. Developmental change in human intestinal alkaline phosphatase. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3909–3912. doi: 10.1073/pnas.75.8.3909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mulivor R. A., Plotkin L. I., Harris H. Differential inhibition of the products of the human alkaline phosphatase loci. Ann Hum Genet. 1978 Jul;42(1):1–13. doi: 10.1111/j.1469-1809.1978.tb00927.x. [DOI] [PubMed] [Google Scholar]
  17. Nathanson L., Fishman W. H. New observations on the Regan isoenzyme of alkaline phosphatase in cancer patients. Cancer. 1971 Jun;27(6):1388–1397. doi: 10.1002/1097-0142(197106)27:6<1388::aid-cncr2820270619>3.0.co;2-0. [DOI] [PubMed] [Google Scholar]
  18. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  19. Sakiyama T., Robinson J. C., Chou J. Y. Characterization of alkaline phosphatases from human first trimester placentas. J Biol Chem. 1979 Feb 10;254(3):935–938. [PubMed] [Google Scholar]
  20. Seargeant L. E., Stinson R. A. Evidence that three structural genes code for human alkaline phosphatases. Nature. 1979 Sep 13;281(5727):152–154. doi: 10.1038/281152a0. [DOI] [PubMed] [Google Scholar]
  21. Singer R. M., Herz F. Multiple effects of glucocorticoids and hyperosmolality on isoenzyme expression in cultured KB cells. J Natl Cancer Inst. 1978 Mar;60(3):573–577. doi: 10.1093/jnci/60.3.573. [DOI] [PubMed] [Google Scholar]
  22. Sussman H. H., Small P. A., Jr, Cotlove E. Human alkaline phosphatase. Immunochemical identification of organ-specific isoenzymes. J Biol Chem. 1968 Jan 10;243(1):160–166. [PubMed] [Google Scholar]
  23. Sussman H. H., Weintraub B. D., Rosen S. W. Relationship of ectopic placental alkaline phosphatase to ectopic chorionic gonadotropin and placental lactogen. Discordance of three "markers" for cancer. Cancer. 1974 Mar;33(3):820–823. doi: 10.1002/1097-0142(197403)33:3<820::aid-cncr2820330329>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  24. Suzuki H., Iino S., Endo Y., Torii M., Miki K. Tumor-specific alkaline phosphatase in hepatoma. Ann N Y Acad Sci. 1975 Aug 22;259:307–320. doi: 10.1111/j.1749-6632.1975.tb25427.x. [DOI] [PubMed] [Google Scholar]
  25. Van Belle H. Kinetics and inhibition of alkaline phosphatases from canine tissues. Biochim Biophys Acta. 1972 Nov 10;289(1):158–168. doi: 10.1016/0005-2744(72)90118-0. [DOI] [PubMed] [Google Scholar]
  26. Wada H. G., Górnicki Z., Sussman H. H. The sialoglycoprotein subunits of human placental brush border membranes characterized by two-two-dimensional electrophoresis. J Supramol Struct. 1977;6(4):473–484. doi: 10.1002/jss.400060402. [DOI] [PubMed] [Google Scholar]
  27. Wada H. G., Shindelman J. E., Ortmeyer A. E., Sussman H. H. Demonstration of placental alkaline phosphatase in human breast cancer. Int J Cancer. 1979 Jun 15;23(6):781–787. doi: 10.1002/ijc.2910230608. [DOI] [PubMed] [Google Scholar]
  28. Whitaker K. B., Moss D. W. Comparison of radioactive peptides obtained from specifically labelled human renal and placental alkaline phosphatases. Biochem J. 1979 Oct 1;183(1):189–192. doi: 10.1042/bj1830189. [DOI] [PMC free article] [PubMed] [Google Scholar]

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