Protein phosphorylation in isolated synaptic junctional structures: changes during development.
Protein phosphorylation in isolated synaptic junctional structures: changes during development.
复制标题
孤立的突触连接结构中的蛋白质磷酸化：发育过程中的变化。
DOI：
10.1016/0006-8993(82)91030-7
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发表时间：
1982
期刊：
影响因子：
2.9
通讯作者：
Kelly,PT
中科院分区：
文献类型：
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作者：
Kelly,PT
在出生后发育的不同阶段，体外研究了突触质膜（SPM）和纯化突触连接（SJ）中富集的亚细胞组分的内源性磷酸化。用内源性和外源性（组蛋白）蛋白作为底物测量蛋白激酶活性。蛋白质磷酸化是由环AMP调节也进行了研究。SPM和SJ组分在出生后5至15天显示出激酶活性和环amp刺激蛋白磷酸化的大量增加。SJ组分在环amp刺激的分子量为73,000 (1a), 68,000 (1b), 65,000 （1c）和55,000的内源性蛋白磷酸化中表现出显著的年龄依赖性变化。用可溶性环amp依赖性激酶磷酸化分离的SJ的实验表明，蛋白1a、1b和1c的磷酸化是由于它们在新生SJ中重新出现，而不是由于新生SJ中已经存在的激酶和底物的成熟依赖性偶联。通过[3H]环AMP结合和光亲和标记[32P]8- n3环AMP来测定突触部分环AMP依赖性激酶的调控亚基水平，并观察到在出生后发育过程中变化不大。这些发现表明，在不同发育阶段分离的SJ片段中，突触的原位外观与内源性、环状amp刺激的蛋白磷酸化的酶促成熟之间存在很强的相关性。
Endogenous phosphorylation in subcellular fractions enriched in synaptic plasma membranes (SPM) and purified synaptic junction (SJ) has been examined in vitro at various stages of postnatal development. Protein kinase activity was measured using both endogenous and exogenous (histones) proteins as substrates. Protein phosphorylation that is regulated by cyclic AMP also was investigated. SPM and SJ fractions displayed large increments in kinase activities and cyclic AMP-stimulated protein phosphorylation between postnatal days 5 and 15. SJ fractions exhibited a dramatic age-dependent change in the cyclic AMP-stimulated phosphorylation of endogenous proteins of molecular weights 73,000 (1a), 68,000 (1b), 65,000 (1c) and 55,000. Experiments in which isolated SJs were phosphorylated with soluble cyclic AMP-dependent kinase showed that the phosphorylation of proteins 1a, 1b and 1c resulted from their de novo appearance in newborn SJ fractions and not from a maturation-dependent coupling of kinases and substrates that were already present in newborn SJ fractions. The levels of regulatory (R) subunits of cyclic AMP-dependent kinases in synaptic fractions were measured by [3H]cyclic AMP binding and photoaffinity labeling with [32P]8-N3-cyclic AMP and were observed to change little during postnatal development. These findings show that there is a strong correlation between the in situ appearance of synapses and the enzymatic maturation of endogenous, cyclic AMP-stimulated protein phosphorylation in SJ fractions isolated at different stages of development.