Identification and characterization of CKLiK, a novel granulocyte Ca(++)/calmodulin-dependent kinase.
Identification and characterization of CKLiK, a novel granulocyte Ca(++)/calmodulin-dependent kinase.
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DOI：
10.1182/blood.v96.9.3215.h8003215_3215_3223
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发表时间：
2000-11
期刊：
影响因子：
20.3
通讯作者：
S. Verploegen;J. Lammers;L. Koenderman;P. Coffer
中科院分区：
文献类型：
--
作者：
S. Verploegen;J. Lammers;L. Koenderman;P. Coffer
人类粒细胞具有多种参与宿主防御的特定效应功能。一些广泛表达的蛋白激酶参与了这些效应器功能的调节。一种基于聚合酶链式反应的策略被用来鉴定新的粒细胞特异性激酶。鉴定了一种新的蛋白激酶互补DNA，其开放阅读框架为357个氨基酸，与钙调素依赖的蛋白激酶I(CaMKI)具有同源性。这被称为CaMKI样激酶(CKLiK)。对造血细胞CKLiK信使RNA(MRNA)表达的分析表明，CKLiK信使RNA在人多形核白细胞(PMN)中几乎是唯一表达的。CD34(+)干细胞在中性粒细胞分化过程中CKLiK基因表达上调。通过体外磷酸化环磷酸腺苷反应元件调节剂(CREM)分析，CKLiK的活性依赖于钙离子和钙调蛋白。此外，与对照细胞相比，离子霉素处理的CKLiK转基因细胞表现出CRE结合蛋白(CREB)转录活性的诱导。此外，CaMK-激动素能增强CKLiK的活性。在体内，通过向稳定表达CKLiK的髓系细胞系中加入白细胞介素8(IL-8)来激活CKLiK。此外，CKLiK的可诱导激活足以诱导细胞外信号相关蛋白(ERK)丝裂原激活蛋白(MAP)的活性。这些数据确定了一种新的依赖于钙(++)/钙调蛋白的PMN特异性激酶，它可能在钙(++)介导的人类粒细胞功能调节中发挥作用。
Human granulocytes are characterized by a variety of specific effector functions involved in host defense. Several widely expressed protein kinases have been implicated in the regulation of these effector functions. A polymerase chain reaction-based strategy was used to identify novel granulocyte-specific kinases. A novel protein kinase complementary DNA with an open reading frame of 357 amino acids was identified with homology to calcium-calmodulin-dependent kinase I (CaMKI). This has been termed CaMKI-like kinase (CKLiK). Analysis of CKLiK messenger RNA (mRNA) expression in hematopoietic cells demonstrated an almost exclusive expression in human polymorphonuclear leukocytes (PMN). Up-regulation of CKLiK mRNA occurs during neutrophilic differentiation of CD34(+) stem cells. CKLiK kinase activity was dependent on Ca(++) and calmodulin as analyzed by in vitro phosphorylation of cyclic adenosine monophosphate responsive element modulator (CREM). Furthermore, CKLiK- transfected cells treated with ionomycin demonstrated an induction of CRE- binding protein (CREB) transcriptional activity compared to control cells. Additionally, CaMK-kinasealpha enhanced CKLiK activity. In vivo activation of CKLiK was shown by addition of interleukin (IL)-8 to a myeloid cell line stably expressing CKLiK. Furthermore inducible activation of CKLiK was sufficient to induce extracellular signal-related kinase (ERK) mitogen-activated protein (MAP) kinase activity. These data identify a novel Ca(++)/calmodulin-dependent PMN- specific kinase that may play a role in Ca(++)-mediated regulation of human granulocyte functions.