Purification and partial characterization of a megakaryocyte colony-stimulating factor from human plasma.
Human plasma obtained from patients with hypomegakaryocytic thrombocytopenia contains a factor that promotes megakaryocyte colony formation by normal human marrow cells. This megakaryocyte colony-stimulating factor was purified from such a plasma specimen. A four-step purification scheme which included ammonium sulfate precipitation, diethylaminoethyl-Sepharose chromatography, affinity chromatography on wheat germ lectin-Sepharose 6MB, and reverse-phase high performance liquid chromatography resulted in a recovery of 16.6% of the initial biological activity and an increase in specific activity by 3,489-fold. The purified protein produced a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified megakaryocyte colony-stimulating factor was capable of promoting megakaryocyte colony formation at a concentration of 7.6 X 10(-8) M. Megakaryocyte colony-stimulating factor was shown to be a glycoprotein and had an apparent 46,000 mol wt. Deglycosylation of megakaryocyte colony-stimulating factor by treatment with trifluoromethane-sulfonate resulted in the loss of its ability to promote megakaryocyte colony formation. Megakaryocyte colony-stimulating factor appears to be an important regulator of in vitro human megakaryocytopoiesis at the level of the colony-forming unit megakaryocyte and may be of importance physiologically.
D003115:Colony-Stimulating Factors / Q000097:blood / Q000302:isolation & purification* / Q000494:pharmacology; D005260:Female; D006410:Hematopoiesis / Q000187:drug effects; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology*; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis; D013921:Thrombocytopenia / Q000097:blood
D003115:Colony-Stimulating Factors; D009844:Oligosaccharides