Protein phosphorlyation in human peripheral blood lymphocytes. Phosphorylation of endogenous plasma membrane and cytoplasmic proteins
- PMID: 228657
- PMCID: PMC1161335
- DOI: 10.1042/bj1820537
Protein phosphorlyation in human peripheral blood lymphocytes. Phosphorylation of endogenous plasma membrane and cytoplasmic proteins
Abstract
Phosphorylation of endogenous proteins in subcellular fractions of human peripheral-blood lymphocytes was studied by one- and two-dimensional polyacrylamide-gel electrophoresis. Studies using extensively purified subcellular fractions indicated that the endogenous phosphorylating activity in the particulate fractions was derived primarily from the plasma membrane. Electrophoresis of (32)P-labelled subcellular fractions in two dimensions [O'Farrell (1975) J. Biol. Chem.250, 4007-4021] provided much greater resolution of the endogenous phosphoproteins than electrophoresis in one dimension, facilitating their excision from gels for quantification of (32)P content. More than 100 cytoplasmic and 20 plasma-membrane phosphorylated species were observed. Phosphorylation of more than 10 cytoplasmic proteins was absolutely dependent on cyclic AMP. In the plasma membrane, cyclic AMP-dependent phosphoproteins were observed with mol.wts. of 42000, 42000, 80000 and 90000 and pI values of 6.1, 6.3, 6.25 and 6.5 respectively. Phosphorylation of endogenous cytoplasmic and plasma-membrane proteins was rapid with t((1/2))=5-12s at 25 degrees C. Between 40 and 70% of the (32)P was recovered as phosphoserine and phosphothreonine when acid hydrolysates of isolated plasma-membrane phosphoproteins were analysed by high-voltage paper electrophoresis. The presence of cyclic AMP-dependent protein kinase and endogenous phosphate-acceptor proteins in the plasma membranes of lymphocytes provides a mechanism by which these cells might respond to plasma-membrane pools of cyclic AMP generated in response to stimulation by mitogens or physiological modulators of lymphocyte function.
Similar articles
-
Protein phosphorylation in human peripheral blood lymphocytes. Subcellular distribution and partial characterization of adenosine 3':5'-cyclic monophosphate-dependent protein kinase.Biochem J. 1979 Aug 15;182(2):525-36. doi: 10.1042/bj1820525. Biochem J. 1979. PMID: 228656 Free PMC article.
-
Phosphorylated lymphocyte plasma-membrane proteins.Biochem J. 1981 Jan 15;194(1):309-18. doi: 10.1042/bj1940309. Biochem J. 1981. PMID: 7305985 Free PMC article.
-
Subcellular distribution of the different platelet proteins phosphorylated on exposure of intact platelets to ionophore A23187 or to prostaglandin E1. Possible role of a membrane phosphopolypeptide in the regulation of calcium-ion transport.Biochem J. 1979 Dec 15;184(3):651-61. doi: 10.1042/bj1840651. Biochem J. 1979. PMID: 120200 Free PMC article.
-
Synaptic membrane proteins as substrates for cyclic AMP-stimulated protein phosphorylation in various regions of rat brain.Biochim Biophys Acta. 1979 Aug 7;555(2):230-8. doi: 10.1016/0005-2736(79)90163-9. Biochim Biophys Acta. 1979. PMID: 224925
-
Protein phosphorylation in rat mast cell granules. Cyclic AMP dependent phosphorylation of a 44K protein associated with broken granules.Biochem Pharmacol. 1987 Jan 1;36(1):131-40. doi: 10.1016/0006-2952(87)90390-x. Biochem Pharmacol. 1987. PMID: 3026401
Cited by
-
Biochemical events associated with lymphocyte activation.Surv Immunol Res. 1984;3(4):295-303. doi: 10.1007/BF02919047. Surv Immunol Res. 1984. PMID: 6095392 Review. No abstract available.
-
Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin.J Cell Biol. 1985 Jul;101(1):207-16. doi: 10.1083/jcb.101.1.207. J Cell Biol. 1985. PMID: 2989299 Free PMC article.
-
Protein phosphorylation in human peripheral blood lymphocytes. Subcellular distribution and partial characterization of adenosine 3':5'-cyclic monophosphate-dependent protein kinase.Biochem J. 1979 Aug 15;182(2):525-36. doi: 10.1042/bj1820525. Biochem J. 1979. PMID: 228656 Free PMC article.
-
Lectins activate lymphocyte pyruvate dehydrogenase by a mechanism sensitive to protease inhibitors.Proc Natl Acad Sci U S A. 1981 Oct;78(10):6256-60. doi: 10.1073/pnas.78.10.6256. Proc Natl Acad Sci U S A. 1981. PMID: 6947229 Free PMC article.
-
Secreted proteins of human monocytes. Analysis by two-dimensional gel electrophoresis and effect of lipopolysaccharide.Biochem J. 1988 Jan 15;249(2):501-11. doi: 10.1042/bj2490501. Biochem J. 1988. PMID: 3257692 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials
Miscellaneous