Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1988 Dec 20;7(13):4163–4167. doi: 10.1002/j.1460-2075.1988.tb03312.x

Phosphorylation of an 85-kd membrane protein by a novel mechanism.

U Seydel 1, W B Huttner 1
PMCID: PMC455127  PMID: 3149582

Abstract

Protein phosphorylation has been recognized as a major mechanism for the regulation of cellular functions. The classical phosphate donor in protein phosphorylation reactions is ATP. Here we show that 3'-phosphoadenosine-5'-phosphosulphate (PAPS), a ubiquitous nucleotide so far known to have a central role in sulphate transfer, serves as phosphate donor for protein phosphorylation. In a very specific, rapid and probably autocatalytic reaction, the 3'-phosphate group of PAPS was found to be transferred to a serine residue of an 85-kd membrane protein (p85). ATP did not serve as phosphate donor in this reaction. Radioactive phosphate incorporated into p85 in a membrane fraction was rapidly lost by dephosphorylation after removal of PAPS or by exchange with unlabelled phosphate after addition of nonradioactive PAPS. PAPS-dependent phosphorylation of the 85-kd protein and other proteins was observed in all rat and bovine tissues examined, as well as in various mammalian cell lines. Our results indicate the existence of a novel widespread form of protein phosphorylation.

Full text

PDF
4163

Images in this article

4163Image
on p.4163
4163Image
on p.4163
4164Image
on p.4164
4164Image
on p.4164
4165Image
on p.4165
4166Image
on p.4166

Selected References

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

  1. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  2. Brandan E., Hirschberg C. B. Purification of rat liver N-heparan-sulfate sulfotransferase. J Biol Chem. 1988 Feb 15;263(5):2417–2422. [PubMed] [Google Scholar]
  3. 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]
  4. Edelman A. M., Blumenthal D. K., Krebs E. G. Protein serine/threonine kinases. Annu Rev Biochem. 1987;56:567–613. doi: 10.1146/annurev.bi.56.070187.003031. [DOI] [PubMed] [Google Scholar]
  5. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  6. Huttner W. B. Tyrosine sulfation and the secretory pathway. Annu Rev Physiol. 1988;50:363–376. doi: 10.1146/annurev.ph.50.030188.002051. [DOI] [PubMed] [Google Scholar]
  7. Khandelwal R. L., Mattoo R. L., Waygood E. B. Phosphoenolpyruvate-dependent protein kinase activity in rat skeletal muscle. FEBS Lett. 1983 Oct 3;162(1):127–132. doi: 10.1016/0014-5793(83)81063-1. [DOI] [PubMed] [Google Scholar]
  8. LIPMANN F. Biological sulfate activation and transfer. Science. 1958 Sep 12;128(3324):575–580. doi: 10.1126/science.128.3324.575. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Lee C., Levin A., Branton D. Copper staining: a five-minute protein stain for sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1987 Nov 1;166(2):308–312. doi: 10.1016/0003-2697(87)90579-3. [DOI] [PubMed] [Google Scholar]
  11. Lee R. W., Huttner W. B. (Glu62, Ala30, Tyr8)n serves as high-affinity substrate for tyrosylprotein sulfotransferase: a Golgi enzyme. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6143–6147. doi: 10.1073/pnas.82.18.6143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lee R. W., Huttner W. B. Tyrosine-O-sulfated proteins of PC12 pheochromocytoma cells and their sulfation by a tyrosylprotein sulfotransferase. J Biol Chem. 1983 Sep 25;258(18):11326–11334. [PubMed] [Google Scholar]
  13. Lee R. W., Suchanek C., Huttner W. B. Direct photoaffinity labeling of proteins with adenosine 3'-[32P]phosphate 5'-phosphosulfate. Atractyloside inhibits labeling of a Mr = 34,000 protein in an adrenal medullary Golgi fraction. J Biol Chem. 1984 Sep 10;259(17):11153–11156. [PubMed] [Google Scholar]
  14. Nairn A. C., Hemmings H. C., Jr, Greengard P. Protein kinases in the brain. Annu Rev Biochem. 1985;54:931–976. doi: 10.1146/annurev.bi.54.070185.004435. [DOI] [PubMed] [Google Scholar]
  15. Renosto F., Seubert P. A., Segel I. H. Adenosine 5'-phosphosulfate kinase from Penicillium chrysogenum. Purification and kinetic characterization. J Biol Chem. 1984 Feb 25;259(4):2113–2123. [PubMed] [Google Scholar]
  16. Schwarz J. K., Capasso J. M., Hirschberg C. B. Translocation of adenosine 3'-phosphate 5'-phosphosulfate into rat liver Golgi vesicles. J Biol Chem. 1984 Mar 25;259(6):3554–3559. [PubMed] [Google Scholar]
  17. Sperling J. Photochemical crosslinking of ATP to histone H4. Photochem Photobiol. 1976 May;23(5):323–326. doi: 10.1111/j.1751-1097.1976.tb07255.x. [DOI] [PubMed] [Google Scholar]
  18. Yue V. T., Schimmel P. R. Direct and specific photochemical cross-linking of adenosine 5'-triphosphate to an aminoacyl-tRNA synthetase. Biochemistry. 1977 Oct 18;16(21):4678–4684. doi: 10.1021/bi00640a023. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES