Cellular Myosin Heavy Chain in Human Platelets : Subcloning of the cDNA Encoding Nonmuscle Myosin II Heavy Chain
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概要
- 論文の詳細を見る
Myosin II, the conventional two-headed myosin that forms bipolar filaments, is directly involved in the regulation of diverse cellular motile processes including cell shape changes and exocytosis of secretory granules during platelet activation. These cellular events are due to cytoskeletal reorganization within the active platelet, which is initiated by actomyosin interactions via complexly orchestrated signalling cascades. Thus, 'actin motor' myosin II plays a crucial role in rearrangement of the platelet cytoskeleton. At least two isoforms of human nonmuscle myosin II heavy chain (MHC), referred to as myosin heavy chain-A and myosin heavy chain-B (MHC-A and MHC-B), have been identified. The entire cDNA sequences which encode these MHC isoforms have been determined. Most tissues express varying ratios of MHC-A and MHC-B, whereas blood platelets express MHC-A exclusively at the protein level. However, previous reports have not shown any direct evidence of mRNA expression of MHC in human platelets. Consequently, the present study investigated endogenous MHC synthesis in human platelets at the transcription level. Peripheral blood platelets were purified from whole blood of a healthy human volunteer by density centrifugation. Total RNA of platelets was extracted using a guanidine-isothiocyanatebased reagent. First-strand cDNA was synthesized from 1 μg of total RNA using 3'-rapid amplification of cDNA ends (3'-RACE), and subjected to PCR amplification of MHC using primers based on the DNA sequences of MHC-A and MHC-B in GenBank. A single amplified PCR product was subcloned into plasmid pBluescript II, and DNA sequencing was performed. Sequences obtained were 100% identical to that registered for MHC-A in GenBank. Moreover, we confirmed by quantitative PCR that blood platelets only express mRNAs encoding MHC-A. We therefore conclude that human platelets express mRNAs which encode MHC-A exclusively, further suggesting that MHC-A is endogenously synthesized in peripheral blood platelets as well as bone marrow megakaryocytes.
- 北里大学の論文
- 2000-08-31
著者
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HIGASHIHARA Masaaki
Department of Hematology, Kitasato University School of Medicine
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Higashihara Masaaki
あけぼの病院 内科
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Higashihara Masaaki
Section Of Hematology Fourth Department Of Internal Medicine Kitasato University School Of Medicine
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Higashihara Masaaki
北里大学 医学部第4内科(血液内科)
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Higashihara Masaaki
Fourth Department Of Internal Medicine School Of Medicine Kitasato University
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Higashihara Masaaki
Department Of Hematology Internal Medicine Kitasato University School Of Medicine
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WATANABE Mariko
Department of Internal Medicine, School of Medicine, Kitasato University
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TAKAISHI Masaaki
Fourth Department of Internal Medicine, School of Medicine, Kitasato University
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Watanabe M
Dep. Of Hematology Internal Medicine Kitasato Univ. School Of Medicine
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Watanabe Mariko
Department Of Anesthesiology Tokai University School Of Medicine
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Takaishi Masaaki
Department of Internal Medicine, Kitasato University School of Medicine
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Takaishi M
Kitasato Univ. School Of Medicine
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