Localization and Biosynthesis of Functional Thrombomodulin in Human Megakaryocytes and a Human Megakaryoblastic Cell Line (MEG-01)

Michinori Ogura; Takahiko Ito; Ikuro Maruyama; Junki Takamatsu; Shuji Yamamoto; Kohei Ogawa; Hiroshi Nagura; Hidehiko Saito

doi:10.1055/s-0038-1647305

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1990; 64(02): 297-301
DOI: 10.1055/s-0038-1647305

Original Article

Schattauer GmbH Stuttgart

Localization and Biosynthesis of Functional Thrombomodulin in Human Megakaryocytes and a Human Megakaryoblastic Cell Line (MEG-01)

Michinori Ogura

¹The First Dept. of Internal Medicine, Nagoya University School of Medicine, Japan

^*The Present address: the Department of Hematology and Cancer Chemotherapeutics, Aichi Cancer Center Hospital, Nagoya, Japan

,

Takahiko Ito

¹The First Dept. of Internal Medicine, Nagoya University School of Medicine, Japan

,

Ikuro Maruyama

²The Third Dept. of Internal Medicine, Kogoshima University School of Medicine, Japan

,

Junki Takamatsu

¹The First Dept. of Internal Medicine, Nagoya University School of Medicine, Japan

,

Shuji Yamamoto

³The Bio-Science Laboratory, Life Science Research Laboratories, Asahi Chemical Industry, Japan

,

Kohei Ogawa

³The Bio-Science Laboratory, Life Science Research Laboratories, Asahi Chemical Industry, Japan

,

Hiroshi Nagura

⁴The Second Dept. of Pathology, Tohoku University School of Medicine, Japan

,

Hidehiko Saito

¹The First Dept. of Internal Medicine, Nagoya University School of Medicine, Japan

› Author Affiliations

Abstract

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Summary

The localization and biosynthesis of functiosnal thrombomodulin (TM) on the cell surfaces of human platelets, megakaryocytes and a human megakaryoblastic cell line (MEG-01) were investigated. TM was demonstrated on the cell surfaces and in cytoplasms of human platelets, megakaryocytes and MEG-01 by an indirect immunofluorescent technique using monospecific rabbit anti-human TM serum. Immunoelectronmicroscopic analysis revealed that TM was localized in plasma membranes of MEG-01 cells as well as human megakaryocytes. ¹²⁵I-monoclonal antithrombomodulin IgG binding assay showed that one MEG-01 cell possessed approximately 78,000 TM molecules on its cell surface. Thrombin-dependent protein C activating-cofactor activity was demonstrated on MEG-01 cells. Northern hybridization technique using cDNA probe of TM revealed that poly(A)⁺-RNA from MEG-01 cells showed a single band of 3.8 kb similar to that from human endothelial cells. These data suggest that human megakaryocytes synthesize functional TM, and thereby platelets possess TM on their surfaces. TM on platelets may participate in the activation of protein C at the site of a hemostatic plug.

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References

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