Sequence analysis of a growth hormone releasing factor from a human pancreatic islet tumor.
Sequence analysis of a growth hormone releasing factor from a human pancreatic islet tumor.
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DOI：
10.1021/bi00267a002
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发表时间：
1982-11
期刊：
影响因子：
2.9
通讯作者：
J. Spiess;J. Rivier;M. Thorner;W. Vale
中科院分区：
文献类型：
--
作者：
J. Spiess;J. Rivier;M. Thorner;W. Vale
纯化肢端肥大症患者的人胰岛肿瘤 (hpGRF) 的生长激素释放因子，并在旋转杯测序仪中进行 Edman 降解。在与异硫氰酸 3-磺基苯酯偶联后或用溴化氰、葡萄球菌蛋白酶或胰蛋白酶裂解后，将约 0.7-1.2 nmol 的肽施加到杯中，无需任何预处理。根据分析数据，确定39个残基的N端序列为H-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-Ser-Arg-Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-。据推测，丙氨酸是残基 40，代表（作为游离酸）hpGRF 的 C 末端。合成的 hpGRF(1-40)-OH 在体外和体内均能高效刺激大鼠垂体前叶分泌 GH。 hpGRF 的 C 端序列似乎对 hpGRF 的生物内在活性和效力没有显着影响，天然产物和合成肽 hpGRF(1-40)-OH、hpGRF(1-40)-NH2 和 hpGRF(1-29)-NH2 显示出相同的体外活性这一事实证明了这一点。根据序列同源性，hpGRF 与胰高血糖素促胰液素家族的成员密切相关，特别是与猪肠道肽 PHI 密切相关。
A growth hormone releasing factor of a human pancreatic islet tumor (hpGRF) of an acromegalic patient was purified and subjected to Edman degradation in a spinning cup sequencer. Approximately 0.7-1.2 nmol of peptide was applied to the cup without any pretreatment, after coupling to 3-sulfophenyl isothiocyanate or after cleavage with cyanogen bromide, staphylococcal protease, or trypsin. On the basis of the analytical data, the N-terminal sequence of 39 residues is established to be H-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-Ser-Arg-Gln-Gln-Gly-Glu-Ser- Asn-Gln-Glu-Arg-Gly-. It is proposed that alanine is residue 40 and represents (as free acid) the C terminus of hpGRF. Synthetic hpGRF(1-40)-OH is highly potent in stimulating GH secretion from the rat anterior pituitary in vitro and in vivo. The C-terminal sequence of hpGRF does not appear to contribute significantly to the biologic intrinsic activity and potency of hpGRF, as demonstrated by the fact that the natural product and the synthetic peptides hpGRF(1-40)-OH, hpGRF(1-40)-NH2, and hpGRF(1-29)-NH2 show equivalent in vitro activities. On the basis of sequence homologies, hpGRF is closely related to members of the glucagon secretin family, especially to the porcine gut peptide PHI.