Purification and Characterization of Thrombopoietin

A thrombopoietic factor, termed thrombopoietin (TPO), was highly purified directly from the plasma of sublethally irradiated 1, 100 rats by measuring the production of megakaryo-cytes from a highly enriched population of rat megakaryocyte progenitor cells (CFU-MK). The rat plasma TPO is a glycoprote...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1995-07, Vol.118 (1), p.229-236
Main Authors: Kato, Takashi, Ogami, Kinya, Shimada, Yoshihiro, Iwamatsu, Akihiro, Sohma, Yoshiaki, Akahori, Hiromichi, Horie, Kaori, Kokubo, Atsuko, Kudo, Yoko, Maeda, Emiko, Kobayashi, Keiko, Ohashi, Hideya, Ozawa, Tadashi, Inoue, Hideo, Kawamura, Kazuo, Miyazaki, Hiroshi
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Biological Assay
Blood Proteins - analysis
c-Mpl ligand
Cell Line
Cloning, Molecular
Colony-Stimulating Factors - analysis
GPI-Linked Proteins
Humans
Key words
Male
megakaryocyte
Megakaryocytes
Membrane Glycoproteins
Mice
Molecular Sequence Data
plasma protein purification
platelet
Proteins - analysis
Rats
Rats, Wistar
Recombinant Proteins - metabolism
throm-bopoietin
Thrombopoietin - chemistry
Thrombopoietin - isolation & purification
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Summary:A thrombopoietic factor, termed thrombopoietin (TPO), was highly purified directly from the plasma of sublethally irradiated 1, 100 rats by measuring the production of megakaryo-cytes 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%108, 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.
ISSN:0021-924X
DOI:10.1093/oxfordjournals.jbchem.a124883