PRIMARY STRUCTURE OF CORTICOTROPIN-RELEASING FACTOR FROM OVINE HYPOTHALAMUS
PRIMARY STRUCTURE OF CORTICOTROPIN-RELEASING FACTOR FROM OVINE HYPOTHALAMUS
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DOI：
10.1073/pnas.78.10.6517
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发表时间：
1981-01-01
期刊：
影响因子：
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通讯作者：
VALE, W
中科院分区：
文献类型：
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作者：
SPIESS, J;RIVIER, J;VALE, W
对绵羊下丘脑41-残基多肽进行了序列分析，该多肽因其高促肾上腺皮质激素释放活性而被假定为促肾上腺皮质激素释放因子（CRF）。在Wittmann-Liebold修饰的Beckman 890C旋转杯测序仪上，利用Edman降解0.6-3.5 nmol的肽段，用反相高压液相色谱法测定了CRF的nh2端39个残基，用于鉴定氨基酸苯基硫代海因（直接微序列分析）。残基40（异亮氨酸）的证据是通过对2.0 nmol乙酰化CRF在分析前被胰蛋白酶选择性切割其精氨酸残基的直接微序列分析提供的。羧基末端热解片段异亮基丙氨酸酰胺被表征为其丹酰衍生物。根据分析数据，我们提出了羊下丘脑CRF的主要结构为：h - ser - gln - glu - pro - pro - ile - ser - leu - asp - leu - thr - phe - his - leu - leu - arg - glu - val - leu - glu - met - thr - lys - ala - asp - gln - leu - gln - gln - gln - his - ser - asn - arg - lys - leu - leu - leu - asp - ile - ala - nh2。与这一建议一致的是，CRF的合成复制在刺激ACTH和β的分泌方面是非常有效的。-endorphin-like immunoactivities。
Sequence analysis was performed of an ovine hypothalamic 41-residue polypeptide that had been postulated to be a putative corticotropin-releasing factor (CRF) because of its high intrinsic ACTH releasing activity. The NH2-terminal 39 residues of CRF were determined by Edman degradation of 0.6-3.5 nmol of peptide in a Wittmann-Liebold modified Beckman 890C spinning cup sequencer with reverse-phase high-pressure liquid chromatography for the identification of amino acid phenylthiohydantoins (direct microsequence analysis). Evidence for residue 40 (isoleucine) was provided by direct microsequence analysis of 2.0 nmol of acetylated CRF selectively cleaved at its arginine residues by trypsin prior to analysis. The thermolytic COOH-terminal fragment isoleucyl-alanineamide was characterized as its dansyl derivative. Based on the analytical data, the following primary structure is proposed for ovine hypothalamic CRF: H-Ser-Gln-Glu-Pro-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. In agreement with this proposal, the synthetic replicate of CRF is highly potent in stimulating secretion of both ACTH and .beta.-endorphin-like immunoactivities.