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. 1986 Oct;83(19):7182–7186. doi: 10.1073/pnas.83.19.7182

Isolation and characterization of a cDNA encoding a human liver/bone/kidney-type alkaline phosphatase.

M J Weiss, P S Henthorn, M A Lafferty, C Slaughter, M Raducha, H Harris
PMCID: PMC386679  PMID: 3532105

Abstract

Alkaline phosphatases (ALPs) [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] isolated from human liver, bone, and kidney (L/B/K) exhibit very similar biochemical and immunologic properties that differentiate them from other human ALPs, such as those characteristically found in placenta and intestine. Despite their similarities, the L/B/K ALPs produced in different tissues show slight physical differences. To examine structural and evolutionary relationships between the various ALPs, a cDNA corresponding to L/B/K ALP mRNA has been isolated. A lambda 11 cDNA expression library was constructed using poly(A) RNA from the osteosarcoma cell line Saos-2 and screened with anti-liver ALP antiserum. The 2553-base-pair cDNA contains an open reading frame that encodes a 524 amino acid polypeptide with a predicted molecular mass of 57.2 kDa. This ALP precursor protein contains a presumed signal peptide of 17 amino acids followed by 37 amino acids that are identical to the amino-terminal sequence determined from purified liver ALP. In addition, amino acid sequences of several CNBr peptides obtained from liver ALP are found within the cDNA-encoded protein. The deduced L/B/K ALP precursor polypeptide shows 52% homology to human placental ALP and 25% homology to Escherichia coli ALP precursor polypeptides. Sixty percent nucleotide homology exists between the human L/B/K and placental cDNAs over the protein coding regions. The 5' and 3' untranslated regions of the L/B/K ALP cDNA, 176 and 805 base pairs, respectively, show no homology to the corresponding regions of placental ALP cDNA.

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Selected References

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

  1. Benham F. J., Fogh J., Harris H. Alkaline phosphatase expression in human cell lines derived from various malignancies. Int J Cancer. 1981 May 15;27(5):637–644. doi: 10.1002/ijc.2910270510. [DOI] [PubMed] [Google Scholar]
  2. Boland C. J., Fried R. M., Tashjian A. H., Jr Measurement of cytosolic free Ca2+ concentrations in human and rat osteosarcoma cells: actions of bone resorption-stimulating hormones. Endocrinology. 1986 Mar;118(3):980–989. doi: 10.1210/endo-118-3-980. [DOI] [PubMed] [Google Scholar]
  3. Boothroyd J. C., Paynter C. A., Cross G. A., Bernards A., Borst P. Variant surface glycoproteins of Trypanosoma brucei are synthesised with cleavable hydrophobic sequences at the carboxy and amino termini. Nucleic Acids Res. 1981 Sep 25;9(18):4735–4743. doi: 10.1093/nar/9.18.4735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradshaw R. A., Cancedda F., Ericsson L. H., Neumann P. A., Piccoli S. P., Schlesinger M. J., Shriefer K., Walsh K. A. Amino acid sequence of Escherichia coli alkaline phosphatase. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3473–3477. doi: 10.1073/pnas.78.6.3473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dale R. M., McClure B. A., Houchins J. P. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. doi: 10.1016/0147-619x(85)90053-8. [DOI] [PubMed] [Google Scholar]
  6. FRASER D. Hypophosphatasia. Am J Med. 1957 May;22(5):730–746. doi: 10.1016/0002-9343(57)90124-9. [DOI] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  8. Garattini E., Hua J. C., Pan Y. C., Udenfriend S. Human liver alkaline phosphatase, purification and partial sequencing: homology with the placental isozyme. Arch Biochem Biophys. 1986 Mar;245(2):331–337. doi: 10.1016/0003-9861(86)90223-7. [DOI] [PubMed] [Google Scholar]
  9. Henthorn P. S., Knoll B. J., Raducha M., Rothblum K. N., Slaughter C., Weiss M., Lafferty M. A., Fischer T., Harris H. Products of two common alleles at the locus for human placental alkaline phosphatase differ by seven amino acids. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5597–5601. doi: 10.1073/pnas.83.15.5597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hua J. C., Berger J., Pan Y. C., Hulmes J. D., Udenfriend S. Partial sequencing of human adult, human fetal, and bovine intestinal alkaline phosphatases: comparison with the human placental and liver isozymes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2368–2372. doi: 10.1073/pnas.83.8.2368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kam W., Clauser E., Kim Y. S., Kan Y. W., Rutter W. J. Cloning, sequencing, and chromosomal localization of human term placental alkaline phosphatase cDNA. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8715–8719. doi: 10.1073/pnas.82.24.8715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kikuchi Y., Yoda K., Yamasaki M., Tamura G. The nucleotide sequence of the promoter and the amino-terminal region of alkaline phosphatase structural gene (phoA) of Escherichia coli. Nucleic Acids Res. 1981 Nov 11;9(21):5671–5678. doi: 10.1093/nar/9.21.5671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kolata G. Novel protein/membrane attachment. Science. 1985 Aug 30;229(4716):850–850. doi: 10.1126/science.229.4716.850. [DOI] [PubMed] [Google Scholar]
  14. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  16. Kraus J. P., Conboy J. G., Rosenberg L. E. Pre-ornithine transcarbamylase. Properties of the cytoplasmic precursor of a mitochondrial matrix enzyme. J Biol Chem. 1981 Nov 10;256(21):10739–10742. [PubMed] [Google Scholar]
  17. Kraus J. P., Rosenberg L. E. Purification of low-abundance messenger RNAs from rat liver by polysome immunoadsorption. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4015–4019. doi: 10.1073/pnas.79.13.4015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Liebhaber S. A., Cash F. E. Locus assignment of alpha-globin structural mutations by hybrid-selected translation. J Clin Invest. 1985 Jan;75(1):64–70. doi: 10.1172/JCI111698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  21. Low M. G., Zilversmit D. B. Role of phosphatidylinositol in attachment of alkaline phosphatase to membranes. Biochemistry. 1980 Aug 19;19(17):3913–3918. doi: 10.1021/bi00558a004. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Meyer L. J., Lafferty M. A., Raducha M. G., Foster C. J., Gogolin K. J., Harris H. Production of a monoclonal antibody to human liver alkaline phosphatase. Clin Chim Acta. 1982 Dec 9;126(2):109–117. doi: 10.1016/0009-8981(82)90026-2. [DOI] [PubMed] [Google Scholar]
  24. Millán J. L. Molecular cloning and sequence analysis of human placental alkaline phosphatase. J Biol Chem. 1986 Mar 5;261(7):3112–3115. [PubMed] [Google Scholar]
  25. Mueller H. D., Stinson R. A., Mohyuddin F., Milne J. K. Isoenzymes of alkaline phosphatase in infantile hypophosphatasia. J Lab Clin Med. 1983 Jul;102(1):24–30. [PubMed] [Google Scholar]
  26. 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]
  27. Robison R. The Possible Significance of Hexosephosphoric Esters in Ossification. Biochem J. 1923;17(2):286–293. doi: 10.1042/bj0170286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Seki T., Chang H. C., Moriuchi T., Denome R., Ploegh H., Silver J. A hydrophobic transmembrane segment at the carboxyl terminus of thy-1. Science. 1985 Feb 8;227(4687):649–651. doi: 10.1126/science.2857501. [DOI] [PubMed] [Google Scholar]
  31. Sowadski J. M., Handschumacher M. D., Murthy H. M., Foster B. A., Wyckoff H. W. Refined structure of alkaline phosphatase from Escherichia coli at 2.8 A resolution. J Mol Biol. 1985 Nov 20;186(2):417–433. doi: 10.1016/0022-2836(85)90115-9. [DOI] [PubMed] [Google Scholar]
  32. Wilbur W. J., Lipman D. J. Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci U S A. 1983 Feb;80(3):726–730. doi: 10.1073/pnas.80.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wyckoff H. W., Handschumacher M., Murthy H. M., Sowadski J. M. The three dimensional structure of alkaline phosphatase from E. coli. Adv Enzymol Relat Areas Mol Biol. 1983;55:453–480. doi: 10.1002/9780470123010.ch6. [DOI] [PubMed] [Google Scholar]

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