Home LiteratureArticle Details
PMID: 2962567 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of calpactin I phospholipid binding by calpactin I light-chain binding and phosphorylation by p60v-src.

The Biochemical journal ·Vol. 247 ·No. 2 ·1987-10-15 ·Pages 321-8

Powell MA, Glenney JR

Abstract

Calpactins I and II are proteins that bind Ca2+, phospholipids, actin and spectrin; they are also major substrates of oncogene and growth-factor-receptor tyrosine kinases. Since calpactins have been proposed to provide a link between membrane lipids and the cytoskeleton, we examined in detail the interactions between purified calpactin I and phospholipid liposomes. We focused on the Ca2+-dependence, the effects of phosphorylation of calpactin I by p60v-src (the protein kinase coded for by the Rous-sarcoma-virus oncogene), and the effects of the binding of calpactin I light chain to calpactin I heavy chain. Binding of the light chain to the heavy chain increased the affinity of calpactin I for phosphatidylserine (PS) liposomes. The opposite effect was observed for phosphorylation by p60v-src; phosphorylation decreased the affinity of calpactin I for PS liposomes. These two opposite effects appeared to be independent, since phosphorylation did not prevent light-chain binding to the heavy chain. Calpactin I was found, by the use of three different techniques, to bind to phospholipid liposomes at less than 10(-8) M free Ca2+. This result is in contrast with those of previous studies, which indicated that greater than 10(-6) M free Ca2+ was required. Our findings suggest that calpactin I may be bound to phospholipids in vivo at Ca2+ concentrations of about 1.5 x 10(-7) M, typical of resting unstimulated cells, and that this interaction may be modulated by light-chain binding and phosphorylation by p60v-src.

MeSH Terms
Annexins Calcium/pharmacology Calcium-Binding Proteins/metabolism Centrifugation, Density Gradient Liposomes/metabolism Oncogene Protein pp60(v-src) Phospholipids/metabolism Phosphorylation Protein Binding/drug effects Protein Kinases/metabolism Retroviridae Proteins/metabolism Tyrosine/metabolism
Chemicals
Annexins Calcium-Binding Proteins Liposomes Phospholipids Retroviridae Proteins Tyrosine Protein Kinases Oncogene Protein pp60(v-src) Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Powell M A
Molecular Biology and Virology Laboratory, Salk Institute for Biological Studies, San Diego, CA 92138.
Glenney J R
References (38)
38 references, click to expand
  1. Transformation by Rous sarcoma virus: effects of src gene expression on the synthesis and phosphorylation of cellular polypeptides.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):5212-6 PMID: 228282
  2. Functionally distinct serine phosphorylation sites of p36, the cellular substrate of retroviral protein kinase; differential inhibition of reassociation with p11.
    EMBO J. 1986 Dec 20;5(13):3455-60 PMID: 2951248
  3. Identification of a cellular protein substrate phosphorylated by the avian sarcoma virus-transforming gene product.
    Cell. 1980 Oct;21(3):829-36 PMID: 6254669
  4. Detection of phosphotyrosine-containing 34,000-dalton protein in the framework of cells transformed with Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1981 Apr;78(4):2388-92 PMID: 6166009
  5. Subcellular location of an abundant substrate (p36) for tyrosine-specific protein kinases.
    Mol Cell Biol. 1983 Mar;3(3):340-50 PMID: 6341813
  6. The 34 kd pp60src substrate is located at the inner face of the plasma membrane.
    Cell. 1983 Jul;33(3):767-79 PMID: 6307526
  7. The p36 substrate of tyrosine-specific protein kinases co-localizes with non-erythrocyte alpha-spectrin antigen, p230, in surface lamina of cultured fibroblasts.
    EMBO J. 1983;2(10):1701-5 PMID: 6357780
  8. Regulation of cell growth and transformation by tyrosine-specific protein kinases: the search for important cellular substrate proteins.
    Curr Top Microbiol Immunol. 1983;107:125-61 PMID: 6421545
  9. Isolation of a calcium-dependent 35-kilodalton substrate for the epidermal growth factor receptor/kinase from A-431 cells.
    J Biol Chem. 1984 Feb 25;259(4):2636-45 PMID: 6698984
  10. Biochemical characterization of a 34-kilodalton normal cellular substrate of pp60v-src and an associated 6-kilodalton protein.
    Mol Cell Biol. 1984 Jan;4(1):77-85 PMID: 6321954
  11. Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin.
    EMBO J. 1984 Jan;3(1):227-33 PMID: 6323166
  12. The role of protein kinase C in cell surface signal transduction and tumour promotion.
    Nature. 1984 Apr 19-25;308(5961):693-8 PMID: 6232463
  13. Calcium-dependent conformational changes in the 36-kDa subunit of intestinal protein I related to the cellular 36-kDa target of Rous sarcoma virus tyrosine kinase.
    J Biol Chem. 1985 Feb 10;260(3):1688-95 PMID: 2981869
  14. Protein-tyrosine kinases.
    Annu Rev Biochem. 1985;54:897-930 PMID: 2992362
  15. Phosphorylation of p36 in vitro with pp60src. Regulation by Ca2+ and phospholipid.
    FEBS Lett. 1985 Nov 11;192(1):79-82 PMID: 2414132
  16. The regulatory chain in the p36-kd substrate complex of viral tyrosine-specific protein kinases is related in sequence to the S-100 protein of glial cells.
    EMBO J. 1985 Nov;4(11):2917-20 PMID: 2998764
  17. Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using Fura-2.
    Nature. 1985 Dec 12-18;318(6046):558-61 PMID: 3934562
  18. Amino-terminal sequence of p36 and associated p10: identification of the site of tyrosine phosphorylation and homology with S-100.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7884-8 PMID: 2415974
  19. The epidermal growth factor-induced calcium signal in A431 cells.
    J Biol Chem. 1986 Jan 5;261(1):279-84 PMID: 3484478
  20. Cloning and expression of human lipocortin, a phospholipase A2 inhibitor with potential anti-inflammatory activity.
    Nature. 1986 Mar 6-12;320(6057):77-81 PMID: 2936963
  21. Binding sites for calcium, lipid and p11 on p36, the substrate of retroviral tyrosine-specific protein kinases.
    FEBS Lett. 1986 Mar 31;198(2):361-4 PMID: 2937654
  22. A consensus amino-acid sequence repeat in Torpedo and mammalian Ca2+-dependent membrane-binding proteins.
    Nature. 1986 Apr 17-23;320(6063):636-8 PMID: 2422556
  23. Protein kinase C activation by diacylglycerol second messengers.
    Cell. 1986 Jun 6;45(5):631-2 PMID: 3708690
  24. Phospholipid-dependent Ca2+ binding by the 36-kDa tyrosine kinase substrate (calpactin) and its 33-kDa core.
    J Biol Chem. 1986 Jun 5;261(16):7247-52 PMID: 2940239
  25. Two related but distinct forms of the Mr 36,000 tyrosine kinase substrate (calpactin) that interact with phospholipid and actin in a Ca2+-dependent manner.
    Proc Natl Acad Sci U S A. 1986 Jun;83(12):4258-62 PMID: 3012561
  26. The cDNA sequence for the protein-tyrosine kinase substrate p36 (calpactin I heavy chain) reveals a multidomain protein with internal repeats.
    Cell. 1986 Jul 18;46(2):201-12 PMID: 3013423
  27. Basal lamina inhibition suppresses synthesis of calcium-dependent proteins associated with mammary epithelial cell spreading.
    Exp Cell Res. 1986 Aug;165(2):450-60 PMID: 3720859
  28. Common domain structure of Ca2+ and lipid-binding proteins.
    FEBS Lett. 1986 Jul 14;203(1):99-103 PMID: 2941317
  29. Association of the S-100-related calpactin I light chain with the NH2-terminal tail of the 36-kDa heavy chain.
    J Biol Chem. 1986 Aug 15;261(23):10485-8 PMID: 2942542
  30. A calcium-dependent 35-kilodalton substrate for epidermal growth factor receptor/kinase isolated from normal tissue.
    J Biol Chem. 1986 Oct 15;261(29):13784-92 PMID: 3020049
  31. Primary structure of bovine calpactin I heavy chain (p36), a major cellular substrate for retroviral protein-tyrosine kinases: homology with the human phospholipase A2 inhibitor lipocortin.
    Biochemistry. 1986 Aug 12;25(16):4497-503 PMID: 2945590
  32. An immunological comparison of several novel calcium-binding proteins.
    J Biol Chem. 1986 Dec 5;261(34):15815-8 PMID: 2946677
  33. The tyrosine phosphorylation substrate p36 is developmentally regulated in embryonic avian limb and is induced in cell culture.
    J Cell Biol. 1986 Nov;103(5):2017-24 PMID: 2430982
  34. The protein-tyrosine kinase substrate p36 is also a substrate for protein kinase C in vitro and in vivo.
    Mol Cell Biol. 1986 Jul;6(7):2738-44 PMID: 2946940
  35. Phosphorylation of lipocortins in vitro by protein kinase C.
    Biochem Biophys Res Commun. 1986 Dec 15;141(2):547-54 PMID: 2948507
  36. Inhibition of phospholipase A2 by "lipocortins" and calpactins. An effect of binding to substrate phospholipids.
    J Biol Chem. 1987 Feb 5;262(4):1698-705 PMID: 2948959
  37. Calpactins: two distinct Ca++-regulated phospholipid- and actin-binding proteins isolated from lung and placenta.
    J Cell Biol. 1987 Mar;104(3):503-11 PMID: 2950118
  38. Calmodulin plays a pivotal role in cellular regulation.
    Science. 1980 Jan 4;207(4426):19-27 PMID: 6243188
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2470321
Published
1987-10-15
Pages
321-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1148411
Subset
IM
Grants
NIGMS NIH HHS · GM 32866 · United States
External Links
PubMed source DOI: 10.1042/bj2470321 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2