A new peptide conjugate as a highly specific substrate for MAP kinase.
A new peptide conjugate as a highly specific substrate for MAP kinase.
复制标题
一种新的肽缀合物，作为 MAP 激酶的高度特异性底物。
DOI：
10.1093/oxfordjournals.jbchem.a021723
复制
发表时间：
1997
期刊：
影响因子：
--
通讯作者：
H. Fujisawa
中科院分区：
文献类型：
--
作者：
I. Kameshita;A. Ishida;H. Fujisawa
合成肽（APRTPGGRC）通过羧基末端半胱氨酰残基与聚-L-赖氨酸交联，被发现是丝裂原活化蛋白（MAP）激酶的高度特异性底物。该肽缀合物表现出比先前用作MAP激酶的特异性底物的游离肽底物（APRTPGGRR，Km > 1 mM）低得多的Km值（74 μ M）。与经常用作MAP激酶底物的髓鞘碱性蛋白不同，这种缀合物不用作cAMP依赖性蛋白激酶、蛋白激酶C或多功能钙调蛋白依赖性蛋白激酶的底物。使用肽缀合物作为底物，MAP激酶活性在粗细胞提取物中直接测定通过体外测定和特异性检测通过凝胶内测定。
A synthetic peptide (APRTPGGRC) cross-linked to poly-L-lysine through a carboxy-terminal cysteinyl residue was found to be a highly specific substrate for mitogen-activated protein (MAP) kinases. This peptide conjugate exhibited a much lower Km value (74 microM) than the free peptide substrate (APRTPGGRR, Km > 1 mM) previously used as a specific substrate for MAP kinases. Unlike myelin basic protein, which has been often used as a substrate for MAP kinases, this conjugate did not serve as substrate for cAMP-dependent protein kinase, protein kinase C, or multifunctional calmodulin-dependent protein kinases. Using the peptide conjugate as a substrate, MAP kinase activities in crude cell extracts were directly determined by in vitro assay and specifically detected by in-gel assay.