Purification and biochemical characterization of human pluripotent hematopoietic colony-stimulating factor.

Autor: Welte, K, Platzer, E, Lu, L, Gabrilove, J L, Levi, E, Mertelsmann, R, Moore, M A
Zdroj: Proceedings of the National Academy of Sciences of the United States of America; March 1985, Vol. 82 Issue: 5 p1526-1530, 5p
Abstrakt: Pluripotent hematopoietic colony-stimulating factor (pluripotent CSF), a protein that is constitutively produced by the human bladder carcinoma cell line 5637, has been purified from low serum (0.2% fetal calf serum)-containing conditioned medium. The purification involved sequential ammonium sulfate precipitation, ion-exchange chromatography, gel filtration, and reversed-phase high-performance liquid chromatography. The purified protein has a molecular weight of 18,000 in NaDodSO4/polyacrylamide gel electrophoresis, both by the silver staining technique and by elution of biological activity from a corresponding gel slice, and has an isoelectric point of 5.5. Pluripotent CSF supports the growth of human mixed colonies, granulocyte-macrophage colonies, and early erythroid colonies and induces differentiation of the human promyelocytic leukemic cell line HL-60 and the murine myelomonocytic leukemic cell line WEHI-3B (D+). The specific activity of the purified pluripotent CSF in the granulocyte-macrophage colony assay is 1.5 X 10(8) units/mg of protein.
Databáze: Supplemental Index