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StCDPK2 的表达和活性揭示了一种高度响应的马铃薯钙依赖蛋白激酶,该激酶参与光信号转导。

StCDPK2 expression and activity reveal a highly responsive potato calcium-dependent protein kinase involved in light signalling.

机构信息

Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), CONICET and Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Vuelta de Obligado 2490 2do piso, 1428, Buenos Aires, Argentina.

出版信息

Planta. 2011 Mar;233(3):593-609. doi: 10.1007/s00425-010-1319-2. Epub 2010 Dec 4.

Abstract

Calcium-dependent protein kinases (CDPKs) are essential calcium sensors. In this work, we have studied StCDPK2 isoform from potato both at gene and protein level. StCdpk2 genomic sequence contains eight exons and seven introns, as was observed for StCdpk1. There is one copy of the gene per genome located in chromosome 7. StCDPK2 encodes an active CDPK of 515 aminoacids, with an apparent MW of 57 kDa, which presents myristoylation and palmitoylation consensus in its N-terminus. StCDPK2 is highly expressed in leaves and green sprouts; enhanced expression was detected under light treatment, which corresponds well with light responsive cis-acting elements found in its promoter sequence. Antibodies against the recombinant StCDPK2::6xHis protein detected this isoform in soluble and particulate fractions from leaves. StCDPK2 autophosphorylation and kinase activity are both calcium dependent reaching half maximal activation at 0.6 μM calcium. The active kinase is autophosphorylated on serine and tyrosine residues and its activity is negatively modulated by phosphatidic acid (PA). Our results reveal StCDPK2 as a signalling element involved in plant growth and development and show that its activity is tightly regulated.

摘要

钙依赖型蛋白激酶(CDPKs)是重要的钙传感器。本研究在基因和蛋白水平上对马铃薯 StCDPK2 同工型进行了研究。StCdpk2 基因组序列包含 8 个外显子和 7 个内含子,与 StCdpk1 相同。每个基因组中都有一个基因位于第 7 号染色体上。StCDPK2 编码一个 515 个氨基酸的活性 CDPK,表观分子量为 57 kDa,其 N 端有豆蔻酰化和棕榈酰化的保守序列。StCDPK2 在叶片和绿芽中高表达;在光照处理下检测到表达增强,与启动子序列中发现的光响应顺式作用元件很好地对应。针对重组 StCDPK2::6xHis 蛋白的抗体在叶片的可溶性和颗粒部分检测到该同工型。StCDPK2 的自磷酸化和激酶活性均依赖于钙,钙达到 0.6 μM 时达到半最大激活。活性激酶在丝氨酸和酪氨酸残基上发生自磷酸化,其活性受到磷酸脂酸(PA)的负调控。我们的结果表明 StCDPK2 是参与植物生长发育的信号元件,并表明其活性受到严格调控。

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