Hanissian S H, Frangakis M, Bland M M, Jawahar S, Chatila T A
Division of Immunology, Children's Hospital, Boston, Massachusetts.
J Biol Chem. 1993 Sep 25;268(27):20055-63.
Ca2+/calmodulin-dependent protein kinase type Gr (CaM kinase-Gr) is a Ca2+/calmodulin-dependent protein kinase which is enriched in the brain and thymus. In this study, we examined the expression of CaM kinase-Gr in human lymphocytes and the regulation of its catalytic activity by antigen receptor signaling. CaM kinase-Gr was found selectively expressed in T lymphocytes in a developmentally regulated manner. It was present at severalfold higher levels in immature thymocytes (CD3low, CD4+CD8+) relative to mature thymocytes (CD3high, CD4+CD8-/CD8+CD4-) or to circulating T lymphocytes. The kinase was preferentially expressed in CD4+ T lymphocytes, but was not detected in B lymphocytes or in monocytes. The impact of T cell antigen receptor-CD3 complex (TCR.CD3) signaling on kinase activity was examined using Jurkat human leukemic T lymphocytes as a model. Treatment of Jurkat cells with anti TCR.CD3 monoclonal antibody induced rapid autophosphorylation of the kinase on serine residues and a dramatic, autophosphorylation-dependent enhancement of both Ca2+/calmodulin-dependent and autonomous kinase activity. Enzyme autophosphorylation and activation were dependent on the influx of extracellular Ca2+ following receptor signaling but could not be induced by an influx of extra-cellular Ca2+ triggered by ionophores, indicating that additional signals delivered via TCR.CD3 contribute to the activation of CaM kinase-Gr. These findings suggest a role for CaM kinase-Gr in T lymphocyte development and activation and indicate the presence of stringent regulatory mechanisms governing the activity of this kinase in situ.
钙/钙调蛋白依赖性蛋白激酶Gr型(CaM激酶-Gr)是一种在脑和胸腺中富集的钙/钙调蛋白依赖性蛋白激酶。在本研究中,我们检测了CaM激酶-Gr在人淋巴细胞中的表达及其催化活性受抗原受体信号传导的调节。发现CaM激酶-Gr以发育调控的方式在T淋巴细胞中选择性表达。相对于成熟胸腺细胞(CD3高,CD4 + CD8 - / CD8 + CD4 - )或循环T淋巴细胞,其在未成熟胸腺细胞(CD3低,CD4 + CD8 +)中的水平高出几倍。该激酶在CD4 + T淋巴细胞中优先表达,但在B淋巴细胞或单核细胞中未检测到。以Jurkat人白血病T淋巴细胞为模型,研究了T细胞抗原受体-CD3复合物(TCR.CD3)信号传导对激酶活性的影响。用抗TCR.CD3单克隆抗体处理Jurkat细胞可诱导该激酶在丝氨酸残基上快速自磷酸化,并显著增强钙/钙调蛋白依赖性和自主激酶活性,且这种增强依赖于自磷酸化。酶的自磷酸化和激活依赖于受体信号传导后细胞外Ca2 +的流入,但不能由离子载体触发的细胞外Ca2 +流入诱导,这表明通过TCR.CD3传递的其他信号有助于CaM激酶-Gr的激活。这些发现提示CaM激酶-Gr在T淋巴细胞发育和激活中起作用,并表明存在严格的调控机制来原位控制该激酶的活性。