Purification and characterization of thrombopoietin.
Purification and characterization of thrombopoietin.
复制标题
血小板生成素的纯化和表征。
DOI：
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
Hiroshi Miyazaki
中科院分区：
文献类型：
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作者：
T. Kato;K. Ogami;Y. Shimada;A. Iwamatsu;Y. Sohma;H. Akahori;K. Horie;Atsuko Kokubo;Yoko Kudo;Emiko Maeda;K. Kobayashi;H. Ohashi;T. Ozawa;Hideo Inoue;K. Kawamura;Hiroshi Miyazaki
通过测量高度浓缩的大鼠巨核系祖细胞(CFU-MK)的巨核细胞生成，直接从亚致死剂量照射的1100只大鼠的血浆中提纯了一种名为血小板生成素(TPO)的血小板生成因子。大鼠血浆TPO是一种糖蛋白，具有很强的疏水性。总活性约为29%，纯化产率约为1.49×10(8)。对从纯化的19 kDa TPO制备的两个肽片段的氨基酸序列进行了分析，并用于克隆大鼠和人TPO的cDNAs。结果发现，19 kDa的TPO被截短，但至少包含163个氨基酸。人TPO基因的序列分析表明，TPO与c-MPL配体完全相同。在COS-1细胞中表达的大鼠和人TPO在体外对CFU-MK具有显著的活性，并具有刺激小鼠血小板生成的活性。这些结果表明，最初在照射后的大鼠血浆中发现的一种促血小板生成因子是大鼠c-MPL的配体。
A thrombopoietic factor, termed thrombopoietin (TPO), was highly purified directly from the plasma of sublethally irradiated 1,100 rats by measuring the production of megakaryocytes from a highly enriched population of rat megakaryocyte progenitor cells (CFU-MK). The rat plasma TPO is a glycoprotein and strongly hydrophobic. The total activity and purification yields obtained were about 29% and 1.49 x 10(8), respectively. The amino acid sequences of the two peptide fragments prepared from the purified 19 kDa TPO were analyzed, and used for the cloning of rat and human TPO cDNAs. It was found that the 19 kDa TPO was truncated but comprised at least 163 amino acids. The sequence of human TPO cDNA revealed that the TPO was identical to the c-Mpl ligand. Both rat and human TPOs expressed in COS-1 cells exhibited significant activity toward the CFU-MK in vitro, and were active in stimulating platelet production in mice. These results indicate that a thrombopoietic factor originally found in the irradiated rat plasma is a ligand for the rat c-Mpl.