title: Characterization of the truncated thrombopoietin variants.
abstract: Thrombopoietin (Tpo) is a specific cytokine which regulates megakaryocyte differentiation and maturation. We isolated a truncated mouse Tpo cDNA, the product of which turned out to function neither as an active Tpo variant nor as an antagonist. To define the functional domains of the Tpo molecule further, various truncated and point-mutated Tpo molecules were prepared and their biological activity was assayed. It was found that deletion of the amino terminal side of a potential proteolytic cleavage site, Arg-Arg motif, caused complete loss of Tpo's activity, and that point-mutants lacking one of four conserved cysteine residues lost Tpo activity. We also noticed that Tpo activity was inhibited by the reducing agent. Thus, it was concluded that the amino terminal half of the Tpo is sufficient for Tpo activity, and that the cysteine residues, especially the last cysteine residue located two amino acids away from the Arg-Arg motif, are critical for this activity.
authors.affiliation: Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Wada
authors.forename: Y
authors.initials: Y
authors.lastname: Nagata
authors.forename: H
authors.initials: H
authors.lastname: Nagahisa
authors.forename: K
authors.initials: K
authors.lastname: Okutomi
authors.forename: S H
authors.initials: SH
authors.lastname: Ha
authors.forename: T
authors.initials: T
authors.lastname: Ohnuki
authors.forename: T
authors.initials: T
authors.lastname: Kanaya
authors.forename: M
authors.initials: M
authors.lastname: Matsumura
authors.forename: K
authors.initials: K
authors.lastname: Todokoro
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018076:DNA, Complementary; D004229:Dithiothreitol / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D013329:Structure-Activity Relationship; D013926:Thrombopoietin / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D013926:Thrombopoietin; D004229:Dithiothreitol