Purification and characterization of glia maturation factor beta: a growth regulator for neurons and glia.
A protein has been isolated from bovine brains by using a modification of the procedure used to purify glia maturation factor. The method consists of ammonium sulfate precipitation, chromatography with DEAE-Sephacel, Sephadex G-75, and hydroxylapatite columns, passage through a heparin-Sepharose column, and finally fractionation by reverse-phase HPLC with a C4 column. The isolated protein reacts strongly with the mouse monoclonal antibody G2-09 and has a molecular weight of approximately 17,000 and an isoelectric point of pH 4.9. The N terminus is blocked, but tryptic digestion releases 28 peptides, 8 of which have been sequenced. The total known residues add up to more than two-thirds of the entire 140-residue protein, estimated from amino acid composition, and show no sequence homology with any known protein. Reversible thermal renaturation greatly enhances its biological activity. The purified protein stimulates differentiation of normal neurons as well as glial cells. It inhibits the proliferation of the N-18 neuroblastoma line and the C6 glioma line while promoting their phenotypic expression. We designate this protein glia maturation factor beta.
Lim
R
R
Department of Neurology, University of Iowa College of Medicine, Iowa City 52242.
Miller
J F
JF
Zaheer
A
A
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001681:Biological Assay; D002417:Cattle; D002455:Cell Division; D002845:Chromatography; D002942:Circular Dichroism; D005346:Fibroblast Growth Factors / Q000276:immunology; D020904:Glia Maturation Factor; D006133:Growth Substances / Q000276:immunology / Q000302:isolation & purification*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000276:immunology / Q000302:isolation & purification*; D009457:Neuroglia / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D010449:Peptide Mapping; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D020904:Glia Maturation Factor; D006133:Growth Substances; D009419:Nerve Tissue Proteins; D005346:Fibroblast Growth Factors