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. 1988 Mar;85(5):1398–1402. doi: 10.1073/pnas.85.5.1398

Aspartic acid-484 of nascent placental alkaline phosphatase condenses with a phosphatidylinositol glycan to become the carboxyl terminus of the mature enzyme.

R Micanovic 1, C A Bailey 1, L Brink 1, L Gerber 1, Y C Pan 1, J D Hulmes 1, S Udenfriend 1
PMCID: PMC279778  PMID: 3422741

Abstract

A carboxyl-terminal chymotryptic peptide from mature human placental alkaline phosphatase was purified by HPLC and monitored by a specific RIA. Sequencing and amino acid assay showed that the carboxyl terminus of the peptide was aspartic acid, representing residue 484 of the proenzyme as deduced from the corresponding cDNA. Further analysis of the peptide showed it to be a peptidoglycan containing one residue of ethanolamine, one residue of glucosamine, and two residues of neutral hexose. The inositol glycan is apparently linked to the alpha carboxyl group of the aspartic acid through the ethanolamine. Location of the inositol glycan on Asp-484 of the proenzyme indicates that a 29-residue peptide is cleaved from the nascent protein during the post-translational condensation with the phosphatidylinositol-glycan.

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Selected References

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

  1. Bangs J. D., Andrews N. W., Hart G. W., Englund P. T. Posttranslational modification and intracellular transport of a trypanosome variant surface glycoprotein. J Cell Biol. 1986 Jul;103(1):255–263. doi: 10.1083/jcb.103.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berger J., Garattini E., Hua J. C., Udenfriend S. Cloning and sequencing of human intestinal alkaline phosphatase cDNA. Proc Natl Acad Sci U S A. 1987 Feb;84(3):695–698. doi: 10.1073/pnas.84.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Campbell D. G., Gagnon J., Reid K. B., Williams A. F. Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. Biochem J. 1981 Apr 1;195(1):15–30. doi: 10.1042/bj1950015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cross G. A. Eukaryotic protein modification and membrane attachment via phosphatidylinositol. Cell. 1987 Jan 30;48(2):179–181. doi: 10.1016/0092-8674(87)90419-3. [DOI] [PubMed] [Google Scholar]
  6. Ezra E., Blacher R., Udenfriend S. Purification and partial sequencing of human placental alkaline phosphatase. Biochem Biophys Res Commun. 1983 Nov 15;116(3):1076–1083. doi: 10.1016/s0006-291x(83)80252-6. [DOI] [PubMed] [Google Scholar]
  7. Fatemi S. H., Tartakoff A. M. Hydrophilic anchor-deficient Thy-1 is secreted by a class E mutant T lymphoma. Cell. 1986 Aug 29;46(5):653–657. doi: 10.1016/0092-8674(86)90340-5. [DOI] [PubMed] [Google Scholar]
  8. Ferguson M. A., Duszenko M., Lamont G. S., Overath P., Cross G. A. Biosynthesis of Trypanosoma brucei variant surface glycoproteins. N-glycosylation and addition of a phosphatidylinositol membrane anchor. J Biol Chem. 1986 Jan 5;261(1):356–362. [PubMed] [Google Scholar]
  9. Ferguson M. A., Low M. G., Cross G. A. Glycosyl-sn-1,2-dimyristylphosphatidylinositol is covalently linked to Trypanosoma brucei variant surface glycoprotein. J Biol Chem. 1985 Nov 25;260(27):14547–14555. [PubMed] [Google Scholar]
  10. Harris H. Multilocus enzyme systems and the evolution of gene expression: the alkaline phosphatases as a model example. Harvey Lect. 1980;76:95–124. [PubMed] [Google Scholar]
  11. Hawke D., Yuan P. M., Shively J. E. Microsequence analysis of peptides and proteins. II. Separation of amino acid phenylthiohydantoin derivatives by high-performance liquid chromatography on octadecylsilane supports. Anal Biochem. 1982 Mar 1;120(2):302–311. doi: 10.1016/0003-2697(82)90351-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  14. Honda S., Takahashi M., Kakehi K., Ganno S. Rapid, automated analysis of monosaccharides by high-performance anion-exchange chromatography of borate complexes with fluorimetric detection using 2-cyanoacetamide. Anal Biochem. 1981 May 1;113(1):130–138. doi: 10.1016/0003-2697(81)90055-5. [DOI] [PubMed] [Google Scholar]
  15. Howard A. D., Berger J., Gerber L., Familletti P., Udenfriend S. Characterization of the phosphatidylinositol-glycan membrane anchor of human placental alkaline phosphatase. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6055–6059. doi: 10.1073/pnas.84.17.6055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ikezawa H., Yamanegi M., Taguchi R., Miyashita T., Ohyabu T. Studies on phosphatidylinositol phosphodiesterase (phospholipase C type) of Bacillus cereus. I. purification, properties and phosphatase-releasing activity. Biochim Biophys Acta. 1976 Nov 19;450(2):154–164. [PubMed] [Google Scholar]
  17. Jemmerson R., Low M. G. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase. Biochemistry. 1987 Sep 8;26(18):5703–5709. doi: 10.1021/bi00392a019. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Kitagawa T., Aikawa T. Enzyme coupled immunoassay of insulin using a novel coupling reagent. J Biochem. 1976 Jan;79(1):233–236. doi: 10.1093/oxfordjournals.jbchem.a131053. [DOI] [PubMed] [Google Scholar]
  20. Krakow J. L., Hereld D., Bangs J. D., Hart G. W., Englund P. T. Identification of a glycolipid precursor of the Trypanosoma brucei variant surface glycoprotein. J Biol Chem. 1986 Sep 15;261(26):12147–12153. [PubMed] [Google Scholar]
  21. Low M. G. Biochemistry of the glycosyl-phosphatidylinositol membrane protein anchors. Biochem J. 1987 May 15;244(1):1–13. doi: 10.1042/bj2440001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Low M. G., Finean J. B. Release of alkaline phosphatase from membranes by a phosphatidylinositol-specific phospholipase C. Biochem J. 1977 Oct 1;167(1):281–284. doi: 10.1042/bj1670281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Low M. G., Kincade P. W. Phosphatidylinositol is the membrane-anchoring domain of the Thy-1 glycoprotein. Nature. 1985 Nov 7;318(6041):62–64. doi: 10.1038/318062a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Ovitt C. E., Strauss A. W., Alpers D. H., Chou J. Y., Boime I. Expression of different-sized placental alkaline phosphatase mRNAs in placenta and choriocarcinoma cells. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3781–3785. doi: 10.1073/pnas.83.11.3781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pan Y. C., Wideman J., Blacher R., Chang M., Stein S. Use of high-performance liquid chromatography for preparing samples for microsequencing. J Chromatogr. 1984 Aug 3;297:13–19. doi: 10.1016/s0021-9673(01)89024-5. [DOI] [PubMed] [Google Scholar]
  28. Slein M. W., Logan G. F. Characterization of the Phospholipases of Bacillus cereus and Their Effects on Erythrocytes, Bone, and Kidney Cells. J Bacteriol. 1965 Jul;90(1):69–81. doi: 10.1128/jb.90.1.69-81.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stein S., Brink L. Amino acid analysis of proteins and peptides at the picomole level: the fluorescamine amino acid analyzer. Methods Enzymol. 1981;79(Pt B):20–25. doi: 10.1016/s0076-6879(81)79008-6. [DOI] [PubMed] [Google Scholar]
  30. Stein S., Moschera J. High-performance liquid chromatography and picomole-level detection of peptides and proteins. Methods Enzymol. 1981;79(Pt B):7–16. doi: 10.1016/s0076-6879(81)79006-2. [DOI] [PubMed] [Google Scholar]
  31. Strang A. M., Williams J. M., Ferguson M. A., Holder A. A., Allen A. K. Trypanosoma brucei brucei variant surface glycoprotein contains non-N-acetylated glucosamine. Biochem J. 1986 Mar 1;234(2):481–484. doi: 10.1042/bj2340481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tse A. G., Barclay A. N., Watts A., Williams A. F. A glycophospholipid tail at the carboxyl terminus of the Thy-1 glycoprotein of neurons and thymocytes. Science. 1985 Nov 29;230(4729):1003–1008. doi: 10.1126/science.2865810. [DOI] [PubMed] [Google Scholar]
  33. Weiss M. J., Henthorn P. S., Lafferty M. A., Slaughter C., Raducha M., Harris H. Isolation and characterization of a cDNA encoding a human liver/bone/kidney-type alkaline phosphatase. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7182–7186. doi: 10.1073/pnas.83.19.7182. [DOI] [PMC free article] [PubMed] [Google Scholar]

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