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首页> 外文期刊>Journal of Experimental Botany >Isolation, characterization, and localization of AgaSGNH cDNA: a new SGNH-motif plant hydrolase specific to Agave americana L. leaf epidermis
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Isolation, characterization, and localization of AgaSGNH cDNA: a new SGNH-motif plant hydrolase specific to Agave americana L. leaf epidermis

机译：AgaSGNH cDNA的分离，鉴定和定位：一种针对龙舌兰叶表皮的新的SGNH基序植物水解酶

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摘要

GDSL and SGNH hydrolases are lipases involved in a wide range of functions, behaving in many cases as bifunctional enzymes. In this work, the isolation and characterization of AgaSGNH, a cDNA encoding a member of the SGNH-hydrolase superfamily from young leaf epidermis of the monocot Agave americana L., is reported. The protein possesses a typical signal peptide at its N-terminus that allows its secretion to the epidermis cell wall, as verified by immunolocalization experiments. In addition, the AgaSGNH sequence contains a His-Leu-Gly-Ala-Glu (HLGAE) motif which is similar to that observed in other plant acyltransferases. Expression levels by northern blot and in situ localization of the corresponding mRNA, as well as the immunolocalization of the protein in Agave young leaves indicate that the protein is specifically present in the epidermal cells. The detailed study performed in different parts of the Agave leaf confirms two aspects: first, the expression of AgaSGNH is limited to the epidermis, and second, the maximum mRNA levels are found in the epidermis of the youngest zones of the leaf which are especially active in cutin biosynthesis. These levels dramatically decrease in the oldest zone of the leaf, where the presence of AgaSGNH mRNA is undetectable, and the biosynthesis of different cuticle components is severely reduced. These data could be compatible with the hypothesis that AgaSGNH could carry out both the hydrolysis and the transfer, from an activated acyl-CoA to a crescent cutin in Agave americana leaves and, therefore, be involved in the still unknown mechanism of plant cutin biosynthesis.
机译：GDSL和SGNH水解酶是脂肪酶，具有多种功能，在许多情况下表现为双功能酶。在这项工作中，报道了从单子叶植物龙舌兰的幼叶表皮中编码SGNH-水解酶超家族成员的cDNA AgaSGNH的分离和表征。该蛋白在其N端具有典型的信号肽，从而使其分泌到表皮细胞壁上，这已通过免疫定位实验证实。另外，AgaSGNH序列含有His-Leu-Gly-Ala-Glu（HLGAE）基序，其与在其他植物酰基转移酶中观察到的基序相似。通过RNA印迹和相应mRNA的原位定位以及龙舌兰幼叶中蛋白质的免疫定位的表达水平表明该蛋白质特异性地存在于表皮细胞中。在龙舌兰叶片的不同部位进行的详细研究证实了两个方面：首先，AgaSGNH的表达仅限于表皮，其次，在叶片最年轻区域的表皮中发现了最大的mRNA水平。在角质的生物合成中。这些水平在叶片最老的区域急剧下降，在该区域中无法检测到AgaSGNH mRNA的存在，并且不同表皮成分的生物合成也大大减少。这些数据可能与假说AgaSGNH可以进行水解和从龙舌兰叶片中的活化酰基辅酶A到新月角质转移的假设相吻合，因此参与了植物角质生物合成的未知机制。

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