High pressure liquid chromatography purification of UP1 and UP2, two related single-stranded nucleic acid-binding proteins from calf thymus.
Two single-stranded nucleic acid-binding proteins, UP1 and UP2, that were originally reported by Herrick and Alberts (Herrick, G., and Alberts, B. (1976) J. Biol. Chem. 251, 2124-2132) have been purified to apparent homogeneity from calf thymus by high performance liquid chromatography. The amino acid sequence of UP1 (Williams, K. R., Stone, K. L., LoPresti, M. B., Merrill, B. M., and Planck, S. R. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 5666-5670) reveals that UP1 contains 195 amino acids, including one dimethylarginine residue near its COOH terminus. Further analysis of this sequence now demonstrates that UP1 contains a 91-residue internal repeat such that when residues 3-93 (the "A" region) are aligned with residues 94-194 (the "B" region), 32% of the amino acids in these two regions are identical and an additional 39% of those changes that are seen could be accomplished by single base changes. The high degree of internal homology between residues 51-61 and 143-152 and in particular the high density of aromatic and positively charged amino acids in these two regions suggest that residues 51-61 and 143-152 may constitute two independent DNA-binding sites. Solid-phase sequencing of three tryptic peptides that together account for 9% of the 39,500-dalton UP2 protein demonstrate that there is a high degree of sequence homology between UP1 and UP2. Of the 34 residues that have been sequenced in UP2, 44% are identical in both UP1 and UP2. The blocked NH2 terminus, amino acid composition, particularly with regard to its high glycine content and the presence of dimethylarginine, and molecular weight of UP2 suggest this protein is related to proteins that have previously been found associated with heterogeneous RNA. Taken together, these data indicate that both UP1 and UP2 belong to a new family of single-stranded nucleic acid-binding proteins that may be closely related to heterogeneous ribonucleoproteins.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011487:Protein Conformation; D013950:Thymus Gland / Q000032:analysis*; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D004268:DNA-Binding Proteins; D010446:Peptide Fragments; D014357:Trypsin