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 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.
Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., Gunma.
T
T
Kato
K
K
Ogami
Y
Y
Shimada
A
A
Iwamatsu
Y
Y
Sohma
H
H
Akahori
K
K
Horie
A
A
Kokubo
Y
Y
Kudo
E
E
Maeda
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D001798:Blood Proteins / Q000032:analysis*; D002460:Cell Line; D003001:Cloning, Molecular; D003115:Colony-Stimulating Factors / Q000032:analysis*; D058851:GPI-Linked Proteins; D006801:Humans; D008297:Male; D008533:Megakaryocytes; D008562:Membrane Glycoproteins; D000090204:Mesothelin; D051379:Mice; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000378:metabolism; D013926:Thrombopoietin / Q000737:chemistry / Q000302:isolation & purification*
D001798:Blood Proteins; D003115:Colony-Stimulating Factors; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D011506:Proteins; D011994:Recombinant Proteins; D013926:Thrombopoietin; D000090204:Mesothelin