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05.06.2019 | Original Article | Ausgabe 3/2019

International Journal of Hematology 3/2019

Soluble CLEC-2 is generated independently of ADAM10 and is increased in plasma in acute coronary syndrome: comparison with soluble GPVI

Zeitschrift:
International Journal of Hematology > Ausgabe 3/2019
Autoren:
Osamu Inoue, Makoto Osada, Junya Nakamura, Fuminori Kazama, Toshiaki Shirai, Nagaharu Tsukiji, Tomoyuki Sasaki, Hiroshi Yokomichi, Tomotaka Dohi, Makoto Kaneko, Makoto Kurano, Mitsuru Oosawa, Shogo Tamura, Kaneo Satoh, Katsuhiro Takano, Katsumi Miyauchi, Hiroyuki Daida, Yutaka Yatomi, Yukio Ozaki, Katsue Suzuki-Inoue
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1007/​s12185-019-02680-4) contains supplementary material, which is available to authorized users.
Osamu Inoue, Makoto Osada, Junya Nakamura and Fuminori Kazama equally contributed to this work.

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Abstract

Soluble forms of platelet membrane proteins are released upon platelet activation. We previously reported that soluble C-type lectin-like receptor 2 (sCLEC-2) is released as a shed fragment (Shed CLEC-2) or as a whole molecule associated with platelet microparticles (MP-CLEC-2). In contrast, soluble glycoprotein VI (sGPVI) is released as a shed fragment (Shed GPVI), but not as a microparticle-associated form (MP-GPVI). However, mechanism of sCLEC-2 generation or plasma sCLEC-2 has not been fully elucidated. Experiments using metalloproteinase inhibitors/stimulators revealed that ADAM10/17 induce GPVI shedding, but not CLEC-2 shedding, and that shed CLEC-2 was partially generated by MMP-2. Although MP-GPVI was not generated, it was generated in the presence of the ADAM10 inhibitor. Moreover, antibodies against the cytoplasmic or extracellular domain of GPVI revealed the presence of the GPVI cytoplasmic domain, but not the extracellular domain, in the microparticles. These findings suggest that most of the GPVI on microparticles are induced to shed by ADAM10; MP-GPVI is thus undetected. Plasma sCLEC-2 level was 1/32 of plasma sGPVI level in normal subjects, but both soluble proteins significantly increased in plasma of patients with acute coronary syndrome. Thus, sCLEC-2 and sGPVI are released by different mechanisms and released in vivo upon platelet activation.

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Zusatzmaterial
Supplementary material 1 (JPEG 1257 kb)
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Supplementary material 2 (JPEG 1622 kb)
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Supplementary material 3 (JPEG 1738 kb)
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