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Thromb Haemost 1996; 76(06): 1053-1062
DOI: 10.1055/s-0038-1650706
Original Article
Schattauer GmbH Stuttgart

Differential Association of Protein Ser/Thr Phosphatase Types 1 and 2A with the Cytoskeleton upon Platelet Activation

Hideki Toyoda
1   The 2nd Department of Internal Medicine, Mie University School of Medicine, Edobashi, Tsu, Mie, Japan
,
Keiji Nakai
1   The 2nd Department of Internal Medicine, Mie University School of Medicine, Edobashi, Tsu, Mie, Japan
,
Serdar B Omay
1   The 2nd Department of Internal Medicine, Mie University School of Medicine, Edobashi, Tsu, Mie, Japan
,
Hiroshi Shima
2   Carcinogenesis Division, National Cancer Research Institute, Tokyo, Japan
,
Minako Nagao
2   Carcinogenesis Division, National Cancer Research Institute, Tokyo, Japan
,
Hiroshi Shiku
1   The 2nd Department of Internal Medicine, Mie University School of Medicine, Edobashi, Tsu, Mie, Japan
,
Masakatsu Nishikawa
1   The 2nd Department of Internal Medicine, Mie University School of Medicine, Edobashi, Tsu, Mie, Japan
› Author Affiliations
Further Information

Publication History

Received 12 March 1996

Accepted after resubmission 19 August 1996

Publication Date:
11 July 2018 (online)

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Summary

The association of protein Ser/Thr phosphatase type 1(PP1) and type 2A (PP2A) with the cytoskeleton (Triton X-100 insoluble residue) during human platelet activation was investigated. In unstimulated platelets, 40% of total PPl-like activity was present in the Triton-insoluble cytoskeleton, while only 10% of the total PP2A-like activity was present in this fraction. Stimulation with 1 U/ml thrombin produced a 1.8-fold increase in PPl-like activity and a 7-fold increase in PP2A-like activity, respectively, in the cytoskeletal fraction, under aggregating conditions. Immunoblot analysis revealed that thrombin treatment increased association of PP1 catalytic subunit isozymes (PPlα, PPlβ, PP1γ) and PP2A catalytic subunit with the cytoskeleton, with concomitant decrease of these enzymes in Triton-soluble fractions. The amounts of cytoskeleton-associated PP1 and PP2A depended on the dose of thrombin which could activate platelets. Agonist-induced redistribution of PP1 and PP2A into the cytoskeleton was inhibited by OP-41483 (a prostaglandin I2 analog). Interaction of PP2A with cytoskeletal proteins strongly correlates with aggregation, whereas the association of PP1 with cytoskeleton can be detected upon platelet activation, even in the absence of aggregation. Co-extraction of protein kinase C and myosin light chain kinase with the cytoskeleton eventually translocated to the cytoskeleton, but only during aggregation. These results suggest that differential translocation of PP1 and PP2A to the cytoskeleton is involved in platelet activation, and their association with cytoskeletal proteins may regulate phosphorylation levels together with protein kinases in platelets.

 

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