Amino acid sequence of the heat-stable inhibitor of the cAMP-dependent protein kinase from rabbit skeletal muscle.
Amino acid sequence of the heat-stable inhibitor of the cAMP-dependent protein kinase from rabbit skeletal muscle.
复制标题
来自兔骨骼肌的 cAMP 依赖性蛋白激酶热稳定性抑制剂的氨基酸序列。
DOI：
10.1073/pnas.82.17.5732
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发表时间：
1985
影响因子：
11.1
通讯作者：
Krebs,EG
中科院分区：
文献类型：
--
作者：
Scott,JD;Fischer,EH;Takio,K;Demaille,JG;Krebs,EG
用微测序技术测定了兔骨骼肌cAMP依赖性蛋白激酶热稳定抑制剂的氨基酸序列。结构的证明涉及一系列非重叠的胰蛋白酶片段，用于初步鉴定86%的氨基酸。用糜蛋白酶、金黄色葡萄球菌V8蛋白酶和肥大细胞蛋白酶II切割产生的互补片段有助于证明该结构。该抑制剂为75个残基的单链多肽，分子量为7829。它缺乏色氨酸、脯氨酸和含硫氨基酸。抑制剂的氨基末端被未鉴定的基团封闭。该分子的氨基末端区域含有激酶抑制结构域，并且基于残基11-30的序列的合成肽是cAMP依赖性蛋白激酶的有效竞争性抑制剂[Scott，J.D.，费希尔，E. H、Demaille，J. G. & Krebs，E. G.等人（1985）Proc. Acad. Sci. USA 82，4379-4383]。残基14-22显示与cAMP依赖性蛋白激酶的调节亚基的“铰链区”相当同源。分子的其余部分与任何蛋白质的已知氨基酸序列都没有相似性。
The amino acid sequence of rabbit skeletal muscle heat-stable inhibitor of the cAMP-dependent protein kinase has been determined by microsequencing techniques. Proof of the structure involved a series of nonoverlapping tryptic fragments for primary identification of 86% of the amino acids. Complementary fragments generated by cleavage with chymotrypsin, Staphylococcus aureus V8 proteinase, and mast cell proteinase II contributed to proof of the structure. The inhibitor is a single polypeptide chain of 75 residues and has a molecular weight of 7829. It lacks tryptophan, proline, and sulfur-containing amino acids. The amino terminus of the inhibitor is blocked by an unidentified group. The amino-terminal region of the molecule contains the kinase inhibitory domain, and synthetic peptides based on the sequence of residues 11-30 are potent competitive inhibitors of the cAMP-dependent protein kinase [Scott, J. D., Fischer, E. H., Demaille, J. G. & Krebs, E. G. (1985) Proc. Natl. Acad. Sci. USA 82, 4379-4383]. Residues 14-22 show considerable homology to the "hinge-regions" of the regulatory subunits of the cAMP-dependent protein kinase. The remainder of the molecule shows no similarity to the known amino acid sequence of any protein.