Expression of Ca2+/calmodulin-dependent protein kinase types II and IV, and reduced DNA synthesis due to the Ca2+/calmodulin-dependent protein kinase inhibitor KN-62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenyl piperazine) in small cel
Expression of Ca2+/calmodulin-dependent protein kinase types II and IV, and reduced DNA synthesis due to the Ca2+/calmodulin-dependent protein kinase inhibitor KN-62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenyl piperazine) in small cel
复制标题
Ca2/钙调蛋白依赖性蛋白激酶 II 型和 IV 型的表达，以及由于 Ca2/钙调蛋白依赖性蛋白激酶抑制剂 KN-62 (1-[N,O-双(5-异喹啉磺酰基)-N-甲基) 导致的 DNA 合成减少
DOI：
10.1016/s0006-2952(95)02393-3
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发表时间：
1996
影响因子：
5.8
通讯作者：
Porter,RA
中科院分区：
文献类型：
--
作者：
Williams,CL;Phelps,SH;Porter,RA
由于细胞内 Ca2+ 的变化会影响有丝分裂细胞周期的进展，因此我们研究了 Ca2+ 结合蛋白在调节细胞周期进展中的作用。有证据表明，Ca2+/钙调蛋白依赖性蛋白激酶（CaM 激酶）失活可抑制小细胞肺癌 (SCLC) 细胞的细胞周期进展。我们还证明 SCLC 细胞表达 II 型 CaM 激酶 (CaMKII) 和 IV 型 CaM 激酶 (CaMKIV)。五个独立的 SCLC 细胞系表达与 CaMKII β 亚基抗体反应的蛋白质，但没有一个表达可检测到的与 CaMKII α 亚基抗体反应的蛋白质。测试的所有 SCLC 细胞系均表达 CaMKIV 的 (α 和 β 亚型)。SCLC 细胞的 CaMKII 免疫沉淀产生多种在 Ca2+/钙调蛋白存在下自磷酸化的蛋白质。自磷酸化受到 CaMKII(281–302) 肽的抑制，该肽对应于 CaMKII 自抑制结构域，并且通过1-[N,O-双(5-异喹啉磺酰基)-N-甲基-L-酪氨酰基]-4-苯基哌嗪 (KN-62) 是一种特异性 CaM 激酶拮抗剂，Ca2+ 通过电压门控 Ca2+ 通道流入会刺激 SCLC 细胞中 CaMKII 的磷酸化，而 KN-62 会抑制 SCLC 细胞的 DNA 合成。 KN-62 的类似抗增殖作用发生在表达 CaMKII 和 CaMKIV 的 SK-N-SH 人神经母细胞瘤细胞和表达 CaMKII 但不表达 CaMKIV 的 K562 人慢性粒细胞白血病细胞中。SCLC 细胞表达 CaMKII 和 CaMKIV，以及这些细胞对 KN-62 抗增殖作用的敏感性。 CaM 激酶调节 SCLC 增殖。
Because changes in intracellular Ca2+affect progression through the mitotic cell cycle, we investigated the role of Ca2+-binding proteins in regulating cell cycle progression. Evidence was found demonstrating that the inactivation of Ca2+/calmodulin-dependent protein kinase (CaM kinase) inhibits cell cycle progression in small cell lung carcinoma (SCLC) cells. We also demonstrated that SCLC cells express both CaM kinase type II (CaMKII) and CaM kinase type IV (CaMKIV). Five independent SCLC cell lines expressed proteins reactive with antibody to the CaMKII β subunit, but none expressed detectable proteins reactive with antibody to the CaMKII α subunit. All SCLC cells lines tested expressed both the (α and β isoforms of CaMKIV. Immunoprecipitation of CaMKII from SCLC cells yielded multiple proteins that autophosphorylated in the presence of Ca2+/calmodulin. Autophosphorylation was inhibited by the CaMKII(281–302) peptide, which corresponds to the CaMKII autoinhibitory domain, and by 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62), a specific CaM kinase antagonist. Influx of Ca2+through voltage-gated Ca2+channels stimulated phosphorylation of CaMKII in SCLC cells, and this was inhibited by KN-62. Incubation of SCLC cells with KN-62 potently inhibited DNA synthesis, and slowed progression through S phase. Similar anti-proliferative effects of KN-62 occurred in SK-N-SH human neuroblastoma cells, which express both CaMKII and CaMKIV, and in K562 human chronic myelogenous leukemia cells, which express CaMKII but not CaMKIV. The expression of both CaMKII and CaMKIV by SCLC cells, and the sensitivity of these cells to the anti-proliferative effects of KN-62, suggest a role for CaM kinase in regulating SCLC proliferation.