SPECIFICITY OF PROTEIN-KINASE INHIBITOR PEPTIDES AND INDUCTION OF LONG-TERM POTENTIATION
SPECIFICITY OF PROTEIN-KINASE INHIBITOR PEPTIDES AND INDUCTION OF LONG-TERM POTENTIATION
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DOI：
10.1073/pnas.91.11.4761
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发表时间：
1994-05-24
影响因子：
11.1
通讯作者：
GREENGARD, P
中科院分区：
文献类型：
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作者：
HVALBY, O;HEMMINGS, HC;GREENGARD, P
先前的研究已经使用了合成肽类似物，对应于蛋白激酶C（PKC）的假底物结构域或Ca 2 +/钙调蛋白依赖性蛋白激酶II（CaMKII）的自调节结构域内的序列，试图定义这些蛋白激酶中的每一种对诱导长时程增强（LTP）的贡献。然而，这些抑制剂肽的特异性不是绝对的。利用向大鼠海马CA 1区神经元的细胞内递送，我们已经确定了两种蛋白激酶抑制剂肽PKC-（19-36）和[Ala（286）]CaMKII-（281-302）作为LTP诱导抑制剂的相对效力。两种肽均阻断LTP的诱导;然而，PKC-（19-36）的效力是[Ala（286）] CaMK II-（281-302）的30倍。还测定了PKC-（19-36）、[Ala（286）]CaMKII-（281-302）和几种其它CaMKII肽类似物在体外抑制蛋白激酶的相对特异性。PKC-（19-36）和[Ala（286）] CaMK II-（281-302）在生理学测定中的效力与其体外蛋白激酶抑制的Ki值的比较表明，对于每种肽观察到的LTP诱导的阻断可归因于PKC的抑制。
Previous studies have used synthetic peptide analogs, corresponding to sequences within the pseudosubstrate domain of protein kinase C (PKC) or the autoregulatory domain of Ca2+/calmodulin-dependent protein kinase II (CaMKII), in attempts to define the contribution of each of these protein kinases to induction of long-term potentiation (LTP). However, the specificity of these inhibitor peptides is not absolute. Using intracellular delivery to rat CA1 hippocampal neurons, we have determined the relative potency of two protein kinase inhibitor peptides, PKC-(19-36) and [Ala(286)]CaMKII-(281-302), as inhibitors of the induction of LTP. Both peptides blocked the induction of LTP; however, PKC-(19-36) was 30-fold more potent than [Ala(286)]CaMKII-(281-302). The relative specificity of PKC-(19-36), [Ala(286)]CaMKII-(281-302), and several other CaMKII peptide analogs for protein kinase inhibition in vitro was also determined. A comparison of the potencies of PKC-(19-36) and [Ala(286)]CaMKII-(281-302) in the physiological assay with their K-i values for protein kinase inhibition in vitro indicates that the blockade of induction of LTP observed for each peptide is attributable to inhibition of PKC.