Pariani Sebastián, Contreras Marisol, Rossi Franco R, Sander Valeria, Corigliano Mariana G, Simón Francisco, Busi María V, Gomez-Casati Diego F, Pieckenstain Fernando L, Duschak Vilma G, Clemente Marina
Unidad de Biotecnología 6-UB6, IIB-INTECH, CONICET-UNSAM, Chascomús, Argentina.
Unidad de Biotecnología 3-UB3, IIB-INTECH, CONICET-UNSAM, Chascomús, Argentina.
Biochimie. 2016 Apr;123:85-94. doi: 10.1016/j.biochi.2016.02.002. Epub 2016 Feb 4.
Many different types of serine proteinase inhibitors have been involved in several kinds of plant physiological processes, including defense mechanisms against phytopathogens. Kazal-type serine proteinase inhibitors, which are included in the serine proteinase inhibitor family, are present in several organisms. These proteins play a regulatory role in processes that involve serine proteinases like trypsin, chymotrypsin, thrombin, elastase and/or subtilisin. In the present work, we characterized two putative Kazal-type serine proteinase inhibitors from Arabidopsis thaliana, which have a single putative Kazal-type domain. The expression of these inhibitors is transiently induced in response to leaf infection by Botrytis cinerea, suggesting that they play some role in defense against pathogens. We also evaluated the inhibitory specificity of one of the Kazal-type serine proteinase inhibitors, which resulted to be induced during the local response to B. cinerea infection. The recombinant Kazal-type serine proteinase inhibitor displayed high specificity for elastase and subtilisin, but low specificity for trypsin, suggesting differences in its selectivity. In addition, this inhibitor exhibited a strong antifungal activity inhibiting the germination rate of B. cinerea conidia in vitro. Due to the important role of proteinase inhibitors in plant protection against pathogens and pests, the information about Kazal-type proteinase inhibitors described in the present work could contribute to improving current methods for plant protection against pathogens.
许多不同类型的丝氨酸蛋白酶抑制剂参与了多种植物生理过程,包括对植物病原体的防御机制。丝氨酸蛋白酶抑制剂家族中的卡扎尔型丝氨酸蛋白酶抑制剂存在于多种生物体中。这些蛋白质在涉及胰蛋白酶、胰凝乳蛋白酶、凝血酶、弹性蛋白酶和/或枯草杆菌蛋白酶等丝氨酸蛋白酶的过程中发挥调节作用。在本研究中,我们对拟南芥中的两种假定的卡扎尔型丝氨酸蛋白酶抑制剂进行了表征,它们具有一个假定的卡扎尔型结构域。这些抑制剂的表达在灰葡萄孢感染叶片后被短暂诱导,表明它们在抵御病原体方面发挥了一定作用。我们还评估了其中一种在对灰葡萄孢感染的局部反应中被诱导的卡扎尔型丝氨酸蛋白酶抑制剂的抑制特异性。重组卡扎尔型丝氨酸蛋白酶抑制剂对弹性蛋白酶和枯草杆菌蛋白酶具有高特异性,但对胰蛋白酶的特异性较低,表明其选择性存在差异。此外,该抑制剂表现出强大的抗真菌活性,可在体外抑制灰葡萄孢分生孢子的萌发率。由于蛋白酶抑制剂在植物抵御病原体和害虫方面的重要作用,本研究中描述的关于卡扎尔型蛋白酶抑制剂的信息可能有助于改进当前的植物病原体防护方法。