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. 1990 Aug 1;269(3):729–734. doi: 10.1042/bj2690729

Purification and characterization of a major phosphatidylserine-binding phosphoprotein from human platelets.

R Burgener 1, M Wolf 1, T Ganz 1, M Baggiolini 1
PMCID: PMC1131648  PMID: 2390065

Abstract

We describe the isolation, lipid-binding properties and partial amino acid sequence of PS-p68, a novel 68 kDa phosphatidylserine-binding protein from human platelets. PS-p68 is an abundant constituent of platelets, accounting for 0.5-0.75% of total cell protein. It was purified from platelet cytosol by affinity chromatography. Amino acid sequence analysis yielded no similarity to identified proteins. In contrast with most known phospholipid-binding proteins, PS-p68 does not bind Ca2+ and does not require Ca2+ for its binding of phosphatidylserine. Phosphatidylserine binding to PS-p68 was inhibited by phosphatidic acid and by alkylphospholipids. PS-p68 was isolated as a major phosphoprotein from 32P-labelled platelets and was found to function as a protein kinase C substrate in vitro. However, treatment of intact platelets with phorbol 12-myristate 13-acetate, thrombin or carbacyclin did not increase PS-p68 phosphorylation. Platelets appear to be the only blood cells containing PS-p68, which was not detected in neutrophils, monocytes and lymphocytes.

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Selected References

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  1. Clemetson K. J., Bienz D., Zahno M. L., Lüscher E. F. Distribution of platelet glycoproteins and phosphoproteins in hydrophobic and hydrophilic phases in Triton X-114 phase partition. Biochim Biophys Acta. 1984 Dec 19;778(3):463–469. doi: 10.1016/0005-2736(84)90395-x. [DOI] [PubMed] [Google Scholar]
  2. Cooper J. A., Sefton B. M., Hunter T. Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 1983;99:387–402. doi: 10.1016/0076-6879(83)99075-4. [DOI] [PubMed] [Google Scholar]
  3. Crompton M. R., Moss S. E., Crumpton M. J. Diversity in the lipocortin/calpactin family. Cell. 1988 Oct 7;55(1):1–3. doi: 10.1016/0092-8674(88)90002-5. [DOI] [PubMed] [Google Scholar]
  4. Dimitrov D. P., Bühler F. R. Inhibition of adenylate cyclase activity during the reversible shape change in human platelets. Biochem Biophys Res Commun. 1988 Apr 29;152(2):649–657. doi: 10.1016/s0006-291x(88)80088-3. [DOI] [PubMed] [Google Scholar]
  5. Haigler H. T., Fitch J. M., Jones J. M., Schlaepfer D. D. Two lipocortin-like proteins, endonexin II and anchorin CII, may be alternate splices of the same gene. Trends Biochem Sci. 1989 Feb;14(2):48–50. doi: 10.1016/0968-0004(89)90041-8. [DOI] [PubMed] [Google Scholar]
  6. Hunkapiller M. W., Lujan E., Ostrander F., Hood L. E. Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. Methods Enzymol. 1983;91:227–236. doi: 10.1016/s0076-6879(83)91019-4. [DOI] [PubMed] [Google Scholar]
  7. Klee C. B. Ca2+-dependent phospholipid- (and membrane-) binding proteins. Biochemistry. 1988 Sep 6;27(18):6645–6653. doi: 10.1021/bi00418a001. [DOI] [PubMed] [Google Scholar]
  8. Maruyama K., Mikawa T., Ebashi S. Detection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecyl sulfate gel electrophoresis. J Biochem. 1984 Feb;95(2):511–519. doi: 10.1093/oxfordjournals.jbchem.a134633. [DOI] [PubMed] [Google Scholar]
  9. Meers P., Ernst J. D., Düzgünes N., Hong K. L., Fedor J., Goldstein I. M., Papahadjopoulos D. Synexin-like proteins from human polymorphonuclear leukocytes. Identification and characterization of granule-aggregating and membrane-fusing activities. J Biol Chem. 1987 Jun 5;262(16):7850–7858. [PubMed] [Google Scholar]
  10. Morse S. S., Moore P. B. 67 k calcimedin (67 kDa) is distinct from p67 calelectrin and lymphocyte 68 kDa Ca2+-binding protein. Biochem J. 1988 Apr 1;251(1):171–174. doi: 10.1042/bj2510171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  12. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  13. Scott M. G., Crimmins D. L., McCourt D. W., Tarrand J. J., Eyerman M. C., Nahm M. H. A simple in situ cyanogen bromide cleavage method to obtain internal amino acid sequence of proteins electroblotted to polyvinyldifluoride membranes. Biochem Biophys Res Commun. 1988 Sep 30;155(3):1353–1359. doi: 10.1016/s0006-291x(88)81290-7. [DOI] [PubMed] [Google Scholar]
  14. Tous G. I., Fausnaugh J. L., Akinyosoye O., Lackland H., Winter-Cash P., Vitorica F. J., Stein S. Amino acid analysis on polyvinylidene difluoride membranes. Anal Biochem. 1989 May 15;179(1):50–55. doi: 10.1016/0003-2697(89)90198-x. [DOI] [PubMed] [Google Scholar]
  15. Walsh M. J., McDougall J., Wittmann-Liebold B. Extended N-terminal sequencing of proteins of archaebacterial ribosomes blotted from two-dimensional gels onto glass fiber and poly(vinylidene difluoride) membrane. Biochemistry. 1988 Sep 6;27(18):6867–6876. doi: 10.1021/bi00418a032. [DOI] [PubMed] [Google Scholar]
  16. Wolf M., Baggiolini M. Identification of phosphatidylserine-binding proteins in human white blood cells. Biochem J. 1990 Aug 1;269(3):723–728. doi: 10.1042/bj2690723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wolf M., Sahyoun N. Protein kinase C and phosphatidylserine bind to Mr 110,000/115,000 polypeptides enriched in cytoskeletal and postsynaptic density preparations. J Biol Chem. 1986 Oct 5;261(28):13327–13332. [PubMed] [Google Scholar]
  18. Yuen S. W., Chui A. H., Wilson K. J., Yuan P. M. Microanalysis of SDS-PAGE electroblotted proteins. Biotechniques. 1989 Jan;7(1):74–83. [PubMed] [Google Scholar]

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