Esch F, Böhlen P, Ling N, Brazeau P, Guillemin R
Biochem Biophys Res Commun. 1983 Dec 28;117(3):772-9. doi: 10.1016/0006-291x(83)91664-9.
A 44 amino acid peptide with high intrinsic growth hormone releasing activity was isolated from 500 bovine hypothalami by means of acid extraction, immunoaffinity chromatography, gel filtration, and two steps of reverse phase HPLC. The growth hormone releasing factor was structurally characterized by gas phase sequence analysis. Reverse phase liquid chromatography of the native peptide and synthetic replicates showed that the molecule possesses an amide rather than a free acid at its carboxyl terminus. The structure of the peptide was established as: Tyr Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala -Arg-Lys-Leu-Leu-Gln-Asp-Ile-Met-Asn-Arg-Gln-Gln-Gly-Glu-Arg-Asn-Gln -Gly-Ala-Lys-Val-Arg-Leu-NH2 using approximately 2 nmol of material.
通过酸提取、免疫亲和色谱、凝胶过滤以及两步反相高效液相色谱法，从500个牛下丘脑分离出一种具有高内在生长激素释放活性的44个氨基酸的肽。通过气相序列分析对生长激素释放因子进行了结构表征。天然肽和合成复制品的反相液相色谱显示，该分子在其羧基末端具有酰胺而非游离酸。该肽的结构确定为：Tyr Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala -Arg-Lys-Leu-Leu-Gln-Asp-Ile-Met-Asn-Arg-Gln-Gln-Gly-Glu-Arg-Asn-Gln -Gly-Ala-Lys-Val-Arg-Leu-NH2，使用了约2 nmol的材料。
