Isolation and characterization of a cDNA coding cowpea (Vigna unguiculata (L.) Walp.) calcineurin B-like protein-interacting protein kinase, VuCIPK1
Isolation and characterization of a cDNA coding cowpea (Vigna unguiculata (L.) Walp.) calcineurin B-like protein-interacting protein kinase, VuCIPK1
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DOI：
10.5511/plantbiotechnology.25.437
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发表时间：
2008-09
影响因子：
1.6
通讯作者：
M. Imamura;T. Yuasa;Tomoko Takahashi;N. Nakamura;N. Htwe;Shao-hui Zheng;K. Shimazaki;M. Iwaya‐Inoue
中科院分区：
文献类型：
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作者：
M. Imamura;T. Yuasa;Tomoko Takahashi;N. Nakamura;N. Htwe;Shao-hui Zheng;K. Shimazaki;M. Iwaya‐Inoue
我们从豇豆 (Vigna unguiculata (L.) Walp.) 中分离出钙调神经磷酸酶 B 样蛋白 (CBL) 相互作用蛋白激酶 (EC 2.7.11.1) 同源物 (VuCIPK1)，与参与 Ktransport 的 AtCIPK3（86% 氨基酸同一性）显着相似。 RT-PCR检测全株VuCIPK1 mRNA的表达。当豇豆遭受各种环境胁迫时，未检测到 VuCIPK1 mRNA 水平有任何显着变化。使用抗 VuCIPK1 特异性抗体和抗 CBL 抗体进行的免疫印迹测定表明，免疫学上 CIPK 和 CBL 相关的多肽优先与豇豆的膜部分相关。抗VuCIPK1抗体与两种不同的CIPK相关多肽发生交叉反应，根中相对分子质量为51 kDa，成熟叶中相对分子质量为55 kDa。使用豇豆叶提取物进行抗 VuCIPK1 抗体的免疫沉淀测定表明，响应盐胁迫，内源 VuCIPK1 在苏氨酸残基上发生自磷酸化。重组 VuCIPK1 在 Mn 2� 存在下磷酸化酪蛋白。这些观察结果表明膜相关的 VuCIPK1 涉及受其磷酸化状态调节的渗透应激激活激酶。
We isolated a calcineurin B-like protein (CBL)-interacting protein kinase (EC 2.7.11.1) homolog (VuCIPK1) from cowpea (Vigna unguiculata (L.) Walp.) with significant similarity to AtCIPK3 (86% amino acid identity), which is involved in Ktransport. VuCIPK1 mRNA was detected in the whole plant by RT-PCR. Any significant change in VuCIPK1 mRNA levels was not detected when cowpea was subjected to various environmental stresses. Immunoblot assay with an anti-VuCIPK1 specific antibody and an anti-CBL antibody showed that immunologically CIPK- and CBL-related polypeptides were preferentially associated with membrane fractions of cowpea. The anti-VuCIPK1 antibody cross-reacted to two different CIPK-related polypeptides with relative molecular masses of 51 kDa in root and 55 kDa in mature leaf. Immunoprecipitation assay by anti-VuCIPK1 antibody using cowpea leaf extracts showed that endogenous VuCIPK1 was autophosphorylated on the threonine residues in response to salt stress. The recombinant VuCIPK1 phosphorylated casein in the presence of Mn 2� . These observations suggest that membrane-associated VuCIPK1 is involved osmotic stress- activated kinase regulated by its phosphorylation status.