Purification and characterization of cMGF, a novel chicken myelomonocytic growth factor.
We describe the purification of a novel hematopoietic growth factor from conditioned medium of a transformed macrophage cell line. The factor, termed chicken myelomonocytic growth factor (cMGF) stimulates the growth of chicken myeloblasts transformed by myb oncogene-containing retroviruses and induces the formation of macrophage colonies in uninfected chick bone marrow cultures. The biological activity of the factor is destroyed by trypsin and by reducing reagents but not by SDS. Analysis of crude conditioned medium on non-reducing SDS gels reveals two active species of cMGF with mol. wts. of 23 and 27 kd. Incubation of radioiodinated partially purified cMGF with myeloblasts demonstrates the specific binding of 23- and 27-kd components under non-reducing, and 25- and 29-kd components under reducing conditions. Glycosylation inhibition experiments indicate that the larger molecules represent glycosylated forms of a single protein moiety. The 27-kd species has been purified to homogeneity (80 000-fold enrichment) and exerts its half maximal activity at 2 X 10(-12) M and its maximal activity at 3 X 10(-11) M. Antibodies prepared to purified cMGF completely neutralize the growth-stimulating activity of the factor.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002645:Chickens; D006098:Granulocytes / Q000187:drug effects; D006133:Growth Substances / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D006410:Hematopoiesis / Q000187:drug effects; D016298:Hematopoietic Cell Growth Factors; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects; D014357:Trypsin / Q000494:pharmacology
D000906:Antibodies; D006133:Growth Substances; D016298:Hematopoietic Cell Growth Factors; D014357:Trypsin