The role of calpain-myosin 9-Rab7b pathway in mediating the expression of toll-like receptor 4 in platelets

A novel mechanism involved in α-granules trafficking

Jui Chi Tsai, Yi Wen Lin, Chun Yao Huang, Chih Yuan Lin, Yi Ting Tsai, Chun Min Shih, Chung Yi Lee, Yung Hsiang Chen, Chi Yuan Li, Nen Chung Chang, Feng Yen Lin, Chien Sung Tsai

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Toll-like receptors (TLRs) plays a critical role in innate immunity. In 2004, Aslam R. and Shiraki R. first determined that murine and human platelets express functional TLRs. Additionally, Andonegui G. demonstrated that platelets express TLR4, which contributes to thrombocytopenia. However, the underlying mechanisms of TLR4 expression by platelets have been rarely explored until now. The aim of this study was to identify the mechanism of TLR4 expression underlying thrombin treatment. The human washed platelets were used in this study. According to flowcytometry and western blot analysis, the surface levels of TLR4 were significantly enhanced in thrombin-activated human platelets and decreased by TMB-8, calpeptin, and U73122, but not Y27632 (a Rho-associated protein kinase ROCK inhibitor) indicating that thrombin-mediated TLR4 expression was modulated by PAR/PLC pathway, calcium and calpain. Co-immunoprecipitation (co-IP) assay demonstrated that the interaction between TLR4 and myosin-9 (a substrate of calpain) was regulated by calpain; cleavage of myosin-9 enhanced TLR4 expression in thrombin treated platelets. Transmission electron microscope data indicated that human platelets used α-granules to control TLR4 expression; the co-IP experiment suggested that myosin-9 did not coordinate with Rab7b to negatively regulate TLR4 trafficking in thrombin treated platelets. In summary, phospholipase Cγ-calpain-myosin 9-Rab7b axis was responsible for the mechanism underlying the regulation of TLR4 containing α-granules trafficking in thrombin-stimulated platelets, which was involved in coagulation.

Original languageEnglish
Article numbere85833
JournalPLoS One
Volume9
Issue number1
DOIs
Publication statusPublished - Jan 28 2014

Fingerprint

Toll-Like Receptor 4
calpain
Calpain
thrombin
Myosins
myosin
Platelets
granules
Blood Platelets
Thrombin
Toll-Like Receptors
Immunoprecipitation
transmission electron microscopes
phospholipase C
thrombocytopenia
coagulation
protein kinases
Toll-like receptor 4
rho-Associated Kinases
Western blotting

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

The role of calpain-myosin 9-Rab7b pathway in mediating the expression of toll-like receptor 4 in platelets : A novel mechanism involved in α-granules trafficking. / Tsai, Jui Chi; Lin, Yi Wen; Huang, Chun Yao; Lin, Chih Yuan; Tsai, Yi Ting; Shih, Chun Min; Lee, Chung Yi; Chen, Yung Hsiang; Li, Chi Yuan; Chang, Nen Chung; Lin, Feng Yen; Tsai, Chien Sung.

In: PLoS One, Vol. 9, No. 1, e85833, 28.01.2014.

Research output: Contribution to journalArticle

Tsai, Jui Chi ; Lin, Yi Wen ; Huang, Chun Yao ; Lin, Chih Yuan ; Tsai, Yi Ting ; Shih, Chun Min ; Lee, Chung Yi ; Chen, Yung Hsiang ; Li, Chi Yuan ; Chang, Nen Chung ; Lin, Feng Yen ; Tsai, Chien Sung. / The role of calpain-myosin 9-Rab7b pathway in mediating the expression of toll-like receptor 4 in platelets : A novel mechanism involved in α-granules trafficking. In: PLoS One. 2014 ; Vol. 9, No. 1.
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abstract = "Toll-like receptors (TLRs) plays a critical role in innate immunity. In 2004, Aslam R. and Shiraki R. first determined that murine and human platelets express functional TLRs. Additionally, Andonegui G. demonstrated that platelets express TLR4, which contributes to thrombocytopenia. However, the underlying mechanisms of TLR4 expression by platelets have been rarely explored until now. The aim of this study was to identify the mechanism of TLR4 expression underlying thrombin treatment. The human washed platelets were used in this study. According to flowcytometry and western blot analysis, the surface levels of TLR4 were significantly enhanced in thrombin-activated human platelets and decreased by TMB-8, calpeptin, and U73122, but not Y27632 (a Rho-associated protein kinase ROCK inhibitor) indicating that thrombin-mediated TLR4 expression was modulated by PAR/PLC pathway, calcium and calpain. Co-immunoprecipitation (co-IP) assay demonstrated that the interaction between TLR4 and myosin-9 (a substrate of calpain) was regulated by calpain; cleavage of myosin-9 enhanced TLR4 expression in thrombin treated platelets. Transmission electron microscope data indicated that human platelets used α-granules to control TLR4 expression; the co-IP experiment suggested that myosin-9 did not coordinate with Rab7b to negatively regulate TLR4 trafficking in thrombin treated platelets. In summary, phospholipase Cγ-calpain-myosin 9-Rab7b axis was responsible for the mechanism underlying the regulation of TLR4 containing α-granules trafficking in thrombin-stimulated platelets, which was involved in coagulation.",
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AU - Tsai, Yi Ting

AU - Shih, Chun Min

AU - Lee, Chung Yi

AU - Chen, Yung Hsiang

AU - Li, Chi Yuan

AU - Chang, Nen Chung

AU - Lin, Feng Yen

AU - Tsai, Chien Sung

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