Radioactive probes for adrenocorticotropic hormone receptors.
Radioactive probes for adrenocorticotropic hormone receptors.
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促肾上腺皮质激素受体的放射性探针。
DOI：
10.1021/bi00354a023
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Mishra,PK
中科院分区：
文献类型：
--
作者：
Hofmann,K;Romovacek,H;Stehle,CJ;Finn,FM;Bothner-By,AA;Mishra,PK
Department of Chemistry，Pittsburgh，Pennsylvania 15213 Received May 28，1985; Revised Mandarin pt Received October 18，1985摘要：我们试图开发可用于ACTH受体鉴定和分离的促肾上腺皮质激素（ACTH）类似物，首先合成含有7Ve-的ACTH片段，在位置25处的（脱硫生物素基）赖氨酸（脱硫生物素）酰胺用于在琼脂糖固定的抗生物素蛋白树脂上亲和色谱纯化酶-受体复合物。因为通过常规碘化技术标记ACTH或ACTH片段由于Met 4的氧化和碘掺入Tyr 2而破坏生物活性，所以我们制备了[Phe 2，Nle 4] ACTH 1_24，[Phe 2，Nle 4，biocy-tin 25] ACTH！25酰胺和[Phe 2，Nle 4，dethiobiocytin 25] ACTH 25酰胺的合成，并对产物进行了HPLC分析和氨基酸分析，结果表明这些产物纯度较高。通过NMR评估肽中Trp残基的叔丁基化的量，发现其小于0.5%。所有三种肽在刺激牛肾上腺皮质细胞中类固醇生成和cAMP形成的能力方面与标准ACTH β等效。[Phe 2，Nle 4] ACTH的碘化！_24，用碘原作为氧化剂，已经完成，没有任何可检测的生物活性损失。[Phe 2，Nle 4] ACTH的单碘和二碘衍生物！_24进行了制备、HPLC分离和生物活性测定。两种肽都具有刺激牛肾上腺皮质细胞中类固醇生成和cAMP产生的全部能力。
Department of Chemistry, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213 Received May 28, 1985; Revised Manuscript Received October 18, 1985 abstract: Our attemptsto develop adrenocorticotropic hormone (ACTH) analogues that can be employed for ACTH receptor identification and isolation began with the synthesis of ACTH fragments containing 7Ve-(dethiobiotinyl) lysine (dethiobiocytin) amide in position 25 to be used for affinity chromatographic purification of hormone-receptor complexes on Sepharose-immobilized avidin resins. Because labeling ACTH or ACTH fragments by conventional iodination techniques destroys biological activity due to oxidation of Met4 and incorporation of iodine into Tyr2, we have prepared [Phe2, Nle4] ACTH1_24,[Phe2, Nle4, biocy-tin25] ACTH! _25 amide, and [Phe2, Nle4, dethiobiocytin25] ACTH^ s amide by conventional synthetic techniques.The HPLC profiles and amino acid analyses of the final products indicatethat the materials are of a high degree of purity. The amount of tertiary butylation of the Trp residue in the peptides was assessed by NMR and was found to be less than 0.5%. All three peptides are equipotent with the standard ACTH^ as concerns their ability to stimulate steroidogenesis and cAMP formation in bovine adrenal cortical cells. Iodination of [Phe2, Nle4] ACTH! _24, with iodogen as the oxidizing agent, has been accomplished without any detectable loss of biological activity. The mono-and diiodo derivatives of [Phe2, Nle4] ACTH! _24 havebeen prepared, separated by HPLC, and assayed for biological activity. Both peptides have the full capacity to stimulate steroidogenesis and cAMP production in bovine adrenal cortical cells.