Molecular cloning and amino acid sequence of the precursor form of bovine adrenodoxin: evidence for a previously unidentified COOH-terminal peptide.
Molecular cloning and amino acid sequence of the precursor form of bovine adrenodoxin: evidence for a previously unidentified COOH-terminal peptide.
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牛肾上腺素还蛋白前体形式的分子克隆和氨基酸序列：先前未识别的 COOH 末端肽的证据。
DOI：
10.1073/pnas.82.17.5705
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发表时间：
1985
影响因子：
11.1
通讯作者：
Waterman,MR
中科院分区：
文献类型：
--
作者：
Okamura,T;John,ME;Zuber,MX;Simpson,ER;Waterman,MR
已在牛肾上腺皮质 cDNA 文库中鉴定出几种对线粒体铁硫蛋白肾上腺氧还蛋白具有特异性的重组 cDNA 克隆。一个克隆 (pBAdx4) 含有 900 个碱基对的插入片段，其中包括肾上腺氧还蛋白前体蛋白的整个氨基酸编码区。从pBAdx4的核苷酸序列推导的成熟肾上腺素的氨基酸序列与通过蛋白质测序确定的氨基酸序列相同，除了三个酰胺变化之外。先前未确定的肾上腺氧还蛋白 NH2 末端前体片段（58 个氨基酸）的序列包含几个碱性残基，这是前往线粒体的蛋白质前体片段的特征。此外，成熟肾上腺素序列的 COOH 末端存在 14 个氨基酸的延伸。这是否代表 COOH 末端前体片段尚不清楚。牛肾上腺皮质 RNA 中存在三种不同的肾上腺氧还蛋白 mRNA [1.75、1.4 和 0.95 KB 长]。来自牛黄体、肝脏和肾脏的 RNA 含有与肾上腺氧还蛋白 cDNA 杂交的转录物。在用 ACTH 或二丁酰-cAMP 处理后，培养的牛肾上腺皮质细胞中会发生肾上腺氧还蛋白 mRNA 的积累，类似于在其所服务的线粒体类固醇羟化酶（即胆固醇侧链裂解细胞色素 P-450 和类固醇 11 β-羟化酶细胞色素 P-450）中观察到的情况。
Several recombinant cDNA clones specific for the mitochondrial iron-sulfur protein adrenodoxin have been identified in a bovine adrenocortical cDNA library. One clone (pBAdx4) contains a 900-base-pair insert that includes the entire amino acid coding region of the adrenodoxin precursor protein. The amino acid sequence of mature adrenodoxin deduced from the nucleotide sequence of pBAdx4 is identical with that determined by protein sequencing except for three amide changes. The previously undetermined sequence of the adrenodoxin NH2-terminal precursor segment (58 amino acids) contains several basic residues, a characteristic feature of the precursor segment of proteins destined for mitochondria. In addition, a 14 amino acid extension is present at the COOH terminus of the mature adrenodoxin sequence. Whether this represents a COOH-terminal precursor segment is not clear. Three different adrenodoxin mRNAs are present [1.75, 1.4, and 0.95 kilobase(s) long] in bovine adrenocortical RNA. RNA from bovine corpus luteum, liver, and kidney contains transcripts that hybridize to adrenodoxin cDNA. Accumulation of adrenodoxin mRNA occurs in cultured bovine adrenocortical cells after treatment with ACTH or dibutyryl-cAMP, similar to that observed for the mitochondrial steroid hydroxylases that it services--namely, the cholesterol side-chain-cleavage cytochrome P-450 and the steroid 11 beta-hydroxylase cytochrome P-450.