A synthetic peptidic substrate of minimal size and semioptimal sequence for the protein tyrosine kinase pp60c-src.
A synthetic peptidic substrate of minimal size and semioptimal sequence for the protein tyrosine kinase pp60c-src.
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蛋白酪氨酸激酶 pp60c-src 的最小尺寸和半优化序列的合成肽底物。
DOI：
10.1006/abbi.1996.0048
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Budde,RJ
中科院分区：
文献类型：
--
作者：
Ramdas,L;Obeyesekere,NU;McMurray,JS;Budde,RJ
我们使用了一种新的方法来确定作为蛋白酪氨酸激酶pp60c-src的抑制剂和/或底物的肽的最小尺寸和半最佳序列。酪氨酸周围的优选氨基酸是通过系统研究确定的，其中我们从三肽EYG开始增加了一系列线性肽的长度。使用迭代循环，肽的大小一次增加一个残基，首先在氨基末端，然后在羧基末端。在每个位置合成一系列六种类似物，并作为抑制剂和底物进行测定。氨基酸G、A、L、F、E和K用于半优化每个位置。三肽EYG不是底物，也不是有效的抑制剂。随着肽段大小的增加，Kid从10.0降至0.10 mM，作为底物的最小肽是六肽。从该方法获得的最佳总体肽EFEYAFF具有0.13 mM的aKi值，KmandVmax值分别为0.21 mM和680 nmol/min/mg。发现我们的最佳肽对pp60c-src的底物特异性高于所有其他可生物利用的肽底物。
We used a novel approach to determine the minimal size and semioptimal sequence of a peptide to serve as an inhibitor and/or substrate for the protein tyrosine kinase pp60c-src. The preferred amino acids surrounding tyrosine were determined by a systematic study in which we increased the length of a series of linear peptides starting from the tripeptide EYG. Using an iterative cycle, the size of the peptide was increased one residue at a time, first at the amino terminus and then at the carboxy terminus. A series of six analogs were synthesized at each position and assayed as inhibitors and substrates. The amino acids G, A, L, F, E, and K were used to semioptimize each position. The tripeptide EYG was not a substrate nor an efficient inhibitor. With increasing size of the peptide, theKidecreased from 10.0 to 0.10 mM. The smallest peptide to serve as a substrate was a hexapeptide. The best overall peptide obtained from this method, EFEYAFF, had aKivalue of 0.13 mMwithKmandVmaxvalues of 0.21 mMand 680 nmol/min/mg, respectively. Our best peptide was found to have higher substrate specificity than all other commerically available peptidic substrates for pp60c-src.