Isolation and characterization of the three fractions (DE-I, DE-II and DE-III) of rat-liver Z-protein and the complete primary structure of DE-II.
Three fractions (DE-I, DE-II and DE-III) of Z-protein (fatty acid binding protein) have been isolated from rat liver cytosol by DEAE-cellulose chromatography and characterized. They had the same molecular weight of 14000 and essentially identical amino acid composition. However, compositions of endogenous fatty acids were found to differ strikingly from one another. Long-chain fatty acids detected in DE-II were palmitic, stearic, oleic, linoleic and arachidonic acids. In contrast to DE-II, DE-III contained mainly arachidonic acid. Molar ratios of endogenous long-chain fatty acids to both DE-II and DE-III were estimated to be around 1.0. Unlike the latter two fractions, DE-I was virtually lipid-free. Analyses of the three fractions by polyacrylamide gel electrophoresis, electrofocusing and DEAE-cellulose chromatography before and after delipidation suggested that the difference between DE-I and DE-II was in part due to fatty acids bound to DE-II. In contrast, DE-III appeared to be somewhat different from these forms in its protein structure, though tryptic peptide mappings of the three fractions did not reveal clear differences among them. Analysis of the primary structure was made on the most abundant fraction, DE-II, to investigate the relationship among the three fractions and to other proteins. The protein was a single chain consisting of 127 amino acid residues and had a mostly acetylated NH2 terminus and a free sulfhydryl group. The complete sequence of Z-protein showed striking homology to cellular retinoid binding proteins and peripheral nerve myelin P2 protein, which indicated the presence of a new family of cellular lipid-binding proteins diverged from a common ancestor. A possible intragenic duplication of Z-protein was also suggested.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002352:Carrier Proteins / Q000145:classification / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002621:Chemistry; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids / Q000032:analysis; D008099:Liver / Q000032:analysis*; D010446:Peptide Fragments / Q000302:isolation & purification*; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D013329:Structure-Activity Relationship
D000596:Amino Acids; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D010446:Peptide Fragments