Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1974 Jul;141(1):293–298. doi: 10.1042/bj1410293

Alkaline phosphatase from pig kidney. Microheterogeneity and the role of neuraminic acid

Kunio Hiwada 1,*, Ernst D Wachsmuth 1
PMCID: PMC1168076  PMID: 4455206

Abstract

Several alkaline phosphatases (EC 3.1.3.1) could be obtained from pig kidney brush-border membrane on extraction with butan-1-ol. Three of the multiple forms were separated by DEAE-cellulose chromatography and further purified. They form a regular series with different degrees of glycosylation (mainly owing to N-acetylneuraminic acid), of charge, of molecular weight, of stability to temperature, to pH and to urea, of minimal requirement for Mg2+ and of extractability by butan-1-ol. In contrast, the detectable antigenic sites, the inhibition by amino acids and the pH-dependency of Km and Vmax. were identical for these multiple forms. On treatment with neuraminidase, the multiple forms became identical in all their properties. It was therefore concluded that the microheterogeneity of alkaline phosphatase is due to different degrees of glycosylation at polypeptide chains which appear to be otherwise identical.

Full text

PDF
293

Images in this article

294-1PLATE 1
on p.294-1
294-2PLATE 2
on p.294-2

Selected References

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

  1. Butterworth P. J. Human kidney and urinary alkaline phosphatases. Biochem J. 1968 Apr;107(4):467–472. doi: 10.1042/bj1070467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Butterworth P. J., Moss D. W. Action of neuraminidase on human kidney alkaline phosphatase. Nature. 1966 Feb 19;209(5025):805–806. doi: 10.1038/209805a0. [DOI] [PubMed] [Google Scholar]
  3. ENGSTROM L. Studies on calf-intestinal alkaline phosphatase. I. Chromatographic purification, microheterogeneity and some other properties of the purified enzyme. Biochim Biophys Acta. 1961 Sep 2;52:36–48. doi: 10.1016/0006-3002(61)90901-5. [DOI] [PubMed] [Google Scholar]
  4. Ghosh N. K., Goldman S. S., Fishman W. H. Human placental alkaline phosphatase; a sialoprotein. Enzymologia. 1967 Aug 31;33(2):113–124. [PubMed] [Google Scholar]
  5. Hiwada K., Wachsmuth E. D. Catalytic properties of alkaline phosphatase from pig kidney. Biochem J. 1974 Jul;141(1):283–291. doi: 10.1042/bj1410283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Latner A. L., Parsons M., Skillen A. W. Isoelectric focusing of alkaline phosphatases from human kidney and calf intestine. Enzymologia. 1971 Jan 31;40(1):1–7. [PubMed] [Google Scholar]
  7. ROBINSON J. C., PIERCE J. E. DIFFERENTIAL ACTION OF NEURAMINIDASE ON HUMAN SERUM ALKALINE PHOSPHATASES. Nature. 1964 Oct 31;204:472–473. doi: 10.1038/204472a0. [DOI] [PubMed] [Google Scholar]
  8. Sarawathi S., Bachhawat B. K. Role of neuraminic acid in the heterogeneity of alkaline phosphatase in sheep brain. Biochem J. 1968 Mar;107(2):185–190. doi: 10.1042/bj1070185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Smith J. K., Eaton R. H., Whitby L. G., Moss D. W. Large-scale gel-filtration in the purification of human liver and small intestine alkaline phosphatases. Anal Biochem. 1968 Apr;23(1):84–96. doi: 10.1016/0003-2697(68)90011-0. [DOI] [PubMed] [Google Scholar]
  10. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  11. Wachsmuth E. D., Hiwada K. Alkaline phosphatase from pig kidney. Method of purification and molecular properties. Biochem J. 1974 Jul;141(1):273–282. doi: 10.1042/bj1410273. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES