Isolation and characterization of caprine corticotropin-releasing factor.
The hypophysiotropic peptide, corticotropin-releasing factor (CRF) was isolated from caprine hypothalamic median eminence tissue by means of acid extraction, immunoaffinity chromatography, gel filtration and two steps of reverse-phase high-performance liquid chromatography (RPLC). Amino acid sequence determination using a gas-phase sequencer established the primary structure of the first 39 residues from the NH2-terminus. The nature of the COOH-terminal dipeptide was elucidated by Staphylococcus aureus V8 protease digestion of the native peptide, dansylation of the digest and comparative RPLC studies with the dansylated dipeptides Ile-Ala-NH2, Ile-Ala-OH, Ala-Ile-NH2 and Ala-Ile-OH. The complete structure of the peptide was established as: H-Ser-Gln-Glu-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu- Val-Leu-Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn- Arg-Lys-Leu-Leu-Asp-Ile-Ala-NH2, which is identical to that of ovine CRF.
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003346:Corticotropin-Releasing Hormone / Q000302:isolation & purification*; D003619:Dansyl Compounds; D006041:Goats; D006801:Humans; D008473:Median Eminence / Q000032:analysis*; D013045:Species Specificity
D003619:Dansyl Compounds; D003346:Corticotropin-Releasing Hormone