Purification and properties of rubrophilin: a novel brain specific membrane polypeptide.
Rubrophilin, a unique brain specific polypeptide, was purified to apparent homogeneity from microsomal fractions of bovine brains. The peptide stains pink with Coomassie Brilliant Blue R-250 (C.I. No. 42660) under specific conditions, has an apparent Mr of 53,000, and is acidic with an apparent pI of 4.9. The purification involves initial solubilization of delipidated microsomes in sodium dodecyl sulfate, followed by ammonium sulfate fractionation, reversed ammonium sulfate gradient elution from diatomaceous earth, gel filtration on polyacrylamide (Biogel P-200), gradient elution chromatography from hydroxylapatite, and reverse-phase chromatography from phenyl-Sepharose. A yield of about 5 mg of rubrophilin was obtained from 9 g of microsomal proteins. Amino acid analysis shows that rubrophilin contains only nine amino acids with residues/mol as follows: alanine (102), glutamic acid (97), lysine (65), proline (55), aspartic acid (48), glycine (44), serine (37), threonine (35), and valine (10). Cysteine, methionine, tryptophan, tyrosine, isoleucine, phenylalanine, histidine, and arginine could not be detected. Relative rubrophilin content of vertebrate brains was as follows: mammals greater than birds greater than reptiles greater than fishes. It is present in mouse retina and human neuroblastoma cell cultures but could not be detected in octopus optic lobe or in cultured C-6 rat glioma cells.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002460:Cell Line; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes; D005910:Glioma / Q000032:analysis; D006801:Humans; D007526:Isoelectric Point; D008565:Membrane Proteins / Q000302:isolation & purification*; D051379:Mice; D008861:Microsomes / Q000032:analysis*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D009447:Neuroblastoma / Q000032:analysis; D051381:Rats; D012104:Reptiles; D012394:Rosaniline Dyes
D000596:Amino Acids; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012394:Rosaniline Dyes; C051352:rubrophilin; C024757:Coomassie brilliant blue R