Nelson R B, Linden D J, Hyman C, Pfenninger K H, Routtenberg A
Cresap Neuroscience Laboratory, Northwestern University, Evanston, Illinois 60201.
J Neurosci. 1989 Feb;9(2):381-9. doi: 10.1523/JNEUROSCI.09-02-00381.1989.
Regulation of neural protein kinase C (PKC) activity appears to directly affect the persistence of long-term potentiation (LTP; Akers and Routtenberg, 1985; Lovinger et al., 1985, 1986, 1987; Routtenberg et al., 1985, 1986; Akers et al., 1986; Linden et al., 1987), a model of neural plasticity (Bliss and Lomo, 1973). In addition, the in vitro phosphorylation of a brain-specific PKC substrate, protein F1 (Mr 47 kDa, pl 4.5), has been directly correlated with persistence of LTP (Lovinger et al., 1986). Because PKC has been implicated in neurite outgrowth and is present at high levels in growth cone-rich areas of fetal brain, we investigated and characterized PKC substrates in a preparation of isolated nerve growth cone fragments from fetal rat brain and compared them with PKC substrates found in adult rat hippocampus. Four major proteins in the growth cone preparation showed endogenous phosphorylation levels at least 10-fold greater than any other phosphoproteins. Three of these 4 phosphoproteins, termed pp40, pp46, and pp80 (Katz et al., 1985), were phosphorylated by exogenous PKC in a dose-dependent manner, indicating that PKC activity might be of particular importance relative to other kinases in growth cone function. The 2 most highly labeled PKC substrates, pp46 and pp80, comigrated on 2-dimensional gels with the adult hippocampal proteins F1 and "80k" (Mr 78-80 kDa, pl 4.0), respectively. In addition, similarities in charge heterogeneity, 2-dimensional phosphopeptide maps, and increased phosphorylation in the presence of exogenous PKC or PKC stimulators suggest that protein F1 and 80k are highly homologous to, if not identical to, pp46 and pp80, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
神经蛋白激酶C(PKC)活性的调节似乎直接影响长时程增强(LTP)的持续性(Akers和Routtenberg,1985年;Lovinger等人,1985年、1986年、1987年;Routtenberg等人,1985年、1986年;Akers等人,1986年;Linden等人,1987年),LTP是一种神经可塑性模型(Bliss和Lomo,1973年)。此外,一种脑特异性PKC底物蛋白F1(分子量47 kDa,等电点4.5)的体外磷酸化与LTP的持续性直接相关(Lovinger等人,1986年)。由于PKC与神经突生长有关,且在胎脑富含生长锥的区域含量很高,我们研究并鉴定了从胎鼠脑分离的神经生长锥片段制剂中的PKC底物,并将它们与成年大鼠海马体中的PKC底物进行了比较。生长锥制剂中的四种主要蛋白质显示出内源性磷酸化水平比任何其他磷蛋白至少高10倍。这4种磷蛋白中的3种,称为pp40、pp46和pp80(Katz等人,1985年),被外源性PKC以剂量依赖性方式磷酸化,表明相对于其他激酶,PKC活性在生长锥功能中可能特别重要。两种标记程度最高的PKC底物pp46和pp80,在二维凝胶上分别与成年海马体蛋白F1和“80k”(分子量78 - 80 kDa,等电点4.0)迁移率相同。此外,电荷异质性、二维磷酸肽图谱的相似性,以及在外源性PKC或PKC刺激剂存在下磷酸化增加,表明蛋白F1和80k分别与pp46和pp80高度同源,甚至可能相同。(摘要截短至250字)