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. 2021 Mar 26;5(3):376–389. doi: 10.1002/rth2.12495

Platelet Src family kinases: A tale of reversible phosphorylation

Yotis A Senis 1,✉, Zoltan Nagy 2, Jun Mori 3, Sophia Lane 4, Patrick Lane 4
PMCID: PMC8035799  PMID: 33870023

Abstract

Sarcoma (Src) family kinases (SFKs) have occupied a central place in platelet research for over 40 years. Discovered by virologists and oncologists as the proto proto‐oncogene, Src tyrosine kinase spurred a phenomenal burst of research on reversible tyrosine phosphorylation and signal transduction. For a time, platelets were adopted as the model of choice for studying the biological functions of Src, owing to their ease of isolation, high Src expression, and lack of a nucleus, only to be abandoned due to challenges of culturing and manipulating using common molecular biology‐based techniques. For platelet biologists, SFKs have remained an important area of investigation, initiating and amplifying signals from all major adhesion, activation, and inhibitory receptors, including the integrin αIIbβ3, the collagen receptor complex glycoprotein VI–Fc receptor γ‐chain, the G protein–coupled ADP receptor P2Y12 and the inhibitory receptors platelet endothelial cell adhesion molecule‐1 and G6b‐B. The vital roles of SFKs in platelets is highlighted by the severe phenotypes of null and gain‐of‐function mutations in SFKs in mice and humans, and effects of pharmacologic inhibitors on platelet activation, thrombosis, and hemostasis. The recent description of critical regulators of SFKs in platelets, namely, C‐terminal Src kinase (Csk), Csk homologous kinase (Chk), the receptor‐type protein‐tyrosine phosphatase receptor type J (PTPRJ) helps explain some of the bleeding side effects of tyrosine kinase inhibitors and are novel therapeutic targets for regulating the thrombotic and hemostatic capacity of platelets. Recent findings from Chk, Csk, and PTPRJ knockout mouse models highlighted that SFKs are able to autoinhibit by phosphorylating their C‐terminal tyrosine residues, providing fundamental insights into SFK autoregulation.

Keywords: kinase, phosphatatase, platelets, Src, tyrosine phosphorylation

Essentials.

  • Sarcoma (Src) family kinases (SFKs) are essential for initiating and amplifying platelet activation.

  • Reversible phosphorylation is a primary mode of regulation of SFK activity.

  • The tyrosine kinases C‐terminal Src kinase (Csk) and Csk homologous kinase and phosphatase protein‐tyrosine phosphatase receptor type J are critical regulators of SFKs.

  • Autophosphorylation provides an additional level of SFK regulation.

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AUTHOR CONTRIBUTIONS

YAS developed theme, prepared images, and wrote and revised the manuscript. ZN prepared images and revised the manuscript. JM developed theme and prepared images. SL developed theme, illustration, and design. PL developed theme, illustration, and design.

RELATIONSHIP DISCLOSURE

The authors report no conflicts of interest to disclose.

Acknowledgements

We thank Dr Alexandra Mazharian for critical reading, discussion, and suggestions. ZN was funded by the British Heart Foundation (BHF) and University Hospital Würzburg, and acknowledges support from Prof Bernhard Nieswandt (University of Würzburg); JM was funded by the BHF; and YAS was funded by the BHF, Etablissement Français du Sang, and Inserm.

Handling Editor: Dr Alisa Wolberg.

Funding information

Funding was provided by BHF, Inserm and EFS (YAS). Additional sources of funding can be found in Acknowledgements.

Contributor Information

Yotis A. Senis, Email: yotis.senis@efs.sante.fr, @YotisSenis.

Zoltan Nagy, @ZoltanNagyPhd.

REFERENCES

  • 1. Rous P. A Sarcoma of the fowl transmissible by an agent separable from the tumor cells. J Exp Med. 1911;13(4):397‐411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Fischer EH, Krebs EG. Conversion of phosphorylase b to phosphorylase a in muscle extracts. J Biol Chem. 1955;216(1):121‐132. [PubMed] [Google Scholar]
  • 3. Hanafusa H, Hanafusa T, Rubin H. The defectiveness of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1963;49:572‐580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. Stehelin D, Varmus HE, Bishop JM, Vogt PK. DNA related to the transforming gene(s) of avian sarcoma viruses is present in normal avian DNA. Nature. 1976;260(5547):170‐173. [DOI] [PubMed] [Google Scholar]
  • 5. Stehelin D, Fujita DJ, Padgett T, Varmus HE, Bishop JM. Detection and enumeration of transformation‐defective strains of avian sarcoma virus with molecular hybridization. Virology. 1977;76(2):675‐684. [DOI] [PubMed] [Google Scholar]
  • 6. Oppermann H, Levinson AD, Varmus HE, Levintow L, Bishop JM. Uninfected vertebrate cells contain a protein that is closely related to the product of the avian sarcoma virus transforming gene (src). Proc Natl Acad Sci U S A. 1979;76(4):1804‐1808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Brugge JS, Erikson RL. Identification of a transformation‐specific antigen induced by an avian sarcoma virus. Nature. 1977;269(5626):346‐348. [DOI] [PubMed] [Google Scholar]
  • 8. Collett MS, Erikson RL. Protein kinase activity associated with the avian sarcoma virus Src gene product. Proc Natl Acad Sci U S A. 1978;75(4):2021‐2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Erikson E, Collett MS, Erikson RL. In vitro synthesis of a functional avian sarcoma virus transforming‐gene product. Nature. 1978;274(5674):919‐921. [DOI] [PubMed] [Google Scholar]
  • 10. Eckhart W, Hutchinson MA, Hunter T. An activity phosphorylating tyrosine in polyoma T antigen immunoprecipitates. Cell. 1979;18(4):925‐933. [DOI] [PubMed] [Google Scholar]
  • 11. Hunter T, Sefton BM. Transforming gene product of Rous sarcoma virus phosphorylates tyrosine. Proc Natl Acad Sci U S A. 1980;77(3):1311‐1315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Tonks NK, Diltz CD, Fischer EH. Characterization of the major protein‐tyrosine‐phosphatases of human placenta. J Biol Chem. 1988;263(14):6731‐6737. [PubMed] [Google Scholar]
  • 13. Ostman A, Yang Q, Tonks NK. Expression of DEP‐1, a receptor‐like protein‐tyrosine‐phosphatase, is enhanced with increasing cell density. Proc Natl Acad Sci U S A. 1994;91(21):9680‐9684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. Tonks NK, Charbonneau H, Diltz CD, Fischer EH, Walsh KA. Demonstration that the leukocyte common antigen CD45 is a protein tyrosine phosphatase. Biochemistry. 1988;27(24):8695‐8701. [DOI] [PubMed] [Google Scholar]
  • 15. Zheng XM, Wang Y, Pallen CJ. Cell transformation and activation of pp60c‐src by overexpression of a protein tyrosine phosphatase. Nature. 1992;359(6393):336‐339. [DOI] [PubMed] [Google Scholar]
  • 16. Golden A, Nemeth SP, Brugge JS. Blood platelets express high levels of the pp60c‐src‐specific tyrosine kinase activity. Proc Natl Acad Sci U S A. 1986;83(4):852‐856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Burkhart JM, Vaudel M, Gambaryan S, et al. The first comprehensive and quantitative analysis of human platelet protein composition allows the comparative analysis of structural and functional pathways. Blood. 2012;120(15):e73‐82. [DOI] [PubMed] [Google Scholar]
  • 18. Zeiler M, Moser M, Mann M. Copy number analysis of the murine platelet proteome spanning the complete abundance range. Mol Cell Proteomics. 2014;13(12):3435‐3445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Senis YA, Mazharian A, Mori J. Src family kinases: at the forefront of platelet activation. Blood. 2014;124(13):2013‐2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. Arias‐Salgado EG, Lizano S, Sarkar S, Brugge JS, Ginsberg MH, Shattil SJ. Src kinase activation by direct interaction with the integrin beta cytoplasmic domain. Proc Natl Acad Sci U S A. 2003;100(23):13298‐13302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Obergfell A, Eto K, Mocsai A, et al. Coordinate interactions of Csk, Src, and Syk kinases with [alpha]IIb[beta]3 initiate integrin signaling to the cytoskeleton. J Cell Biol. 2002;157(2):265‐275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. Ezumi Y, Shindoh K, Tsuji M, Takayama H. Physical and functional association of the Src family kinases Fyn and Lyn with the collagen receptor glycoprotein VI‐Fc receptor gamma chain complex on human platelets. J Exp Med. 1998;188(2):267‐276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. Resh MD. Myristylation and palmitylation of Src family members: the fats of the matter. Cell. 1994;76(3):411‐413. [DOI] [PubMed] [Google Scholar]
  • 24. Moran MF, Koch CA, Anderson D, et al. Src homology region 2 domains direct protein‐protein interactions in signal transduction. Proc Natl Acad Sci U S A. 1990;87(21):8622‐8626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Iba H, Cross FR, Garber EA, Hanafusa H. Low level of cellular protein phosphorylation by nontransforming overproduced p60c‐src. Mol Cell Biol. 1985;5(5):1058‐1066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. Cooper JA, Gould KL, Cartwright CA, Hunter T. Tyr527 is phosphorylated in pp60c‐src: implications for regulation. Science. 1986;231(4744):1431‐1434. [DOI] [PubMed] [Google Scholar]
  • 27. Roskoski R Jr. Src kinase regulation by phosphorylation and dephosphorylation. Biochem Biophys Res Commun. 2005;331(1):1‐14. [DOI] [PubMed] [Google Scholar]
  • 28. Xu W, Harrison SC, Eck MJ. Three‐dimensional structure of the tyrosine kinase c‐Src. Nature. 1997;385(6617):595‐602. [DOI] [PubMed] [Google Scholar]
  • 29. Turro E, Greene D, Wijgaerts A, et al. A dominant gain‐of‐function mutation in universal tyrosine kinase SRC causes thrombocytopenia, myelofibrosis, bleeding, and bone pathologies. Sci Transl Med. 2016;8(328):328ra330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30. Okada M, Nakagawa H. A protein tyrosine kinase involved in regulation of pp60c‐src function. J Biol Chem. 1989;264(35):20886‐20893. [PubMed] [Google Scholar]
  • 31. Nada S, Okada M, MacAuley A, Cooper JA, Nakagawa H. Cloning of a complementary DNA for a protein‐tyrosine kinase that specifically phosphorylates a negative regulatory site of p60c‐src. Nature. 1991;351(6321):69‐72. [DOI] [PubMed] [Google Scholar]
  • 32. Nada S, Yagi T, Takeda H, et al. Constitutive activation of Src family kinases in mouse embryos that lack Csk. Cell. 1993;73(6):1125‐1135. [DOI] [PubMed] [Google Scholar]
  • 33. Ellison S, Mori J, Barr AJ, Senis YA. CD148 enhances platelet responsiveness to collagen by maintaining a pool of active Src family kinases. J Thromb Haemost. 2010;8(7):1575‐1583. [DOI] [PubMed] [Google Scholar]
  • 34. Mori J, Wang YJ, Ellison S, et al. Dominant role of the protein‐tyrosine phosphatase CD148 in regulating platelet activation relative to protein‐tyrosine phosphatase‐1B. Arterioscler Thromb Vasc Biol. 2012;32(12):2956‐2965. [DOI] [PubMed] [Google Scholar]
  • 35. Senis YA. Protein‐tyrosine phosphatases: a new frontier in platelet signal transduction. J Thromb Haemost. 2013;11(10):1800‐1813. [DOI] [PubMed] [Google Scholar]
  • 36. Mori J, Nagy Z, Di Nunzio G, et al. Maintenance of murine platelet homeostasis by the kinase Csk and phosphatase CD148. Blood. 2018;131(10):1122‐1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37. Bjorge JD, Pang A, Fujita DJ. Identification of protein‐tyrosine phosphatase 1B as the major tyrosine phosphatase activity capable of dephosphorylating and activating c‐Src in several human breast cancer cell lines. J Biol Chem. 2000;275(52):41439‐41446. [DOI] [PubMed] [Google Scholar]
  • 38. Somani AK, Bignon JS, Mills GB, Siminovitch KA, Branch DR. Src kinase activity is regulated by the SHP‐1 protein‐tyrosine phosphatase. J Biol Chem. 1997;272(34):21113‐21119. [DOI] [PubMed] [Google Scholar]
  • 39. Zhang SQ, Yang W, Kontaridis MI, et al. Shp2 regulates SRC family kinase activity and Ras/Erk activation by controlling Csk recruitment. Mol Cell. 2004;13(3):341‐355. [DOI] [PubMed] [Google Scholar]
  • 40. Gil‐Henn H, Elson A. Tyrosine phosphatase‐epsilon activates Src and supports the transformed phenotype of Neu‐induced mammary tumor cells. J Biol Chem. 2003;278(18):15579‐15586. [DOI] [PubMed] [Google Scholar]
  • 41. Senis YA, Tomlinson MG, Ellison S, et al. The tyrosine phosphatase CD148 is an essential positive regulator of platelet activation and thrombosis. Blood. 2009;113(20):4942‐4954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42. Nagy Z, Mori J, Ivanova VS, Mazharian A, Senis YA. Interplay between the tyrosine kinases Chk and Csk and phosphatase PTPRJ is critical for regulating platelets in mice. Blood. 2020;135(18):1574‐1587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43. Xu C, Gagnon E, Call ME, et al. Regulation of T cell receptor activation by dynamic membrane binding of the CD3epsilon cytoplasmic tyrosine‐based motif. Cell. 2008;135(4):702‐713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44. Nika K, Soldani C, Salek M, et al. Constitutively active Lck kinase in T cells drives antigen receptor signal transduction. Immunity. 2010;32(6):766‐777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45. Thomas JE, Soriano P, Brugge JS. Phosphorylation of c‐Src on tyrosine 527 by another protein tyrosine kinase. Science. 1991;254(5031):568‐571. [DOI] [PubMed] [Google Scholar]
  • 46. Jove R, Kornbluth S, Hanafusa H. Enzymatically inactive p60c‐src mutant with altered ATP‐binding site is fully phosphorylated in its carboxy‐terminal regulatory region. Cell. 1987;50(6):937‐943. [DOI] [PubMed] [Google Scholar]
  • 47. Cooper JA, MacAuley A. Potential positive and negative autoregulation of p60c‐src by intermolecular autophosphorylation. Proc Natl Acad Sci U S A. 1988;85(12):4232‐4236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48. MacAuley A, Okada M, Nada S, Nakagawa H, Cooper JA. Phosphorylation of Src mutants at Tyr 527 in fibroblasts does not correlate with in vitro phosphorylation by CSK. Oncogene. 1993;8(1):117‐124. [PubMed] [Google Scholar]
  • 49. Osusky M, Taylor SJ, Shalloway D. Autophosphorylation of purified c‐Src at its primary negative regulation site. J Biol Chem. 1995;270(43):25729‐25732. [DOI] [PubMed] [Google Scholar]
  • 50. Kim LC, Song L, Haura EB. Src kinases as therapeutic targets for cancer. Nat Rev Clin Oncol. 2009;6(10):587‐595. [DOI] [PubMed] [Google Scholar]
  • 51. Zhang S, Yu D. Targeting Src family kinases in anti‐cancer therapies: turning promise into triumph. Trends Pharmacol Sci. 2012;33(3):122‐128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 52. Roskoski R Jr. Src protein‐tyrosine kinase structure, mechanism, and small molecule inhibitors. Pharmacol Res. 2015;94:9‐25. [DOI] [PubMed] [Google Scholar]
  • 53. Honigberg LA, Smith AM, Sirisawad M, et al. The Bruton tyrosine kinase inhibitor PCI‐32765 blocks B‐cell activation and is efficacious in models of autoimmune disease and B‐cell malignancy. Proc Natl Acad Sci U S A. 2010;107(29):13075‐13080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54. Gratacap MP, Martin V, Valera MC, et al. The new tyrosine‐kinase inhibitor and anticancer drug dasatinib reversibly affects platelet activation in vitro and in vivo. Blood. 2009;114(9):1884‐1892. [DOI] [PubMed] [Google Scholar]
  • 55. Levade M, David E, Garcia C, et al. Ibrutinib treatment affects collagen and von Willebrand factor‐dependent platelet functions. Blood. 2014;124(26):3991‐3995. [DOI] [PubMed] [Google Scholar]
  • 56. Senis YA, Barr AJ. Targeting receptor‐type protein tyrosine phosphatases with biotherapeutics: is outside‐in better than inside‐out? Molecules. 2018;23(3):569. [DOI] [PMC free article] [PubMed] [Google Scholar]

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