Lionetti Vincenzo, Raiola Alessandro, Mattei Benedetta, Bellincampi Daniela
Dipartimento di Biologia e Biotecnologie "C. Darwin", Sapienza Università di Roma, Rome, Italy.
Dipartimento Territorio e Sistemi Agroforestali, Università di Padova, Legnaro (PD), Italy.
PLoS One. 2015 Jul 23;10(7):e0133810. doi: 10.1371/journal.pone.0133810. eCollection 2015.
Pectin is secreted in a highly methylesterified form and partially de-methylesterified in the cell wall by pectin methylesterases (PMEs). PME activity is expressed during plant growth, development and stress responses. PME activity is controlled at the post-transcriptional level by proteins named PME inhibitors (PMEIs). We have identified, expressed and characterized VvPMEI1, a functional PME inhibitor of Vitis vinifera. VvPMEI1 typically affects the activity of plant PMEs and is inactive against microbial PMEs. The kinetics of PMEI-PME interaction, studied by surface plasmon resonance, indicates that the inhibitor strongly interacts with PME at apoplastic pH while the stability of the complex is reduced by increasing the pH. The analysis of VvPMEI1 expression in different grapevine tissues and during grape fruit development suggests that this inhibitor controls PME activity mainly during the earlier phase of berry development. A proteomic analysis performed at this stage indicates a PME isoform as possible target of VvPMEI1.
果胶以高度甲酯化的形式分泌,并在细胞壁中被果胶甲酯酶(PMEs)部分去甲酯化。PME活性在植物生长、发育和应激反应过程中表达。PME活性在转录后水平上受名为PME抑制剂(PMEIs)的蛋白质控制。我们已经鉴定、表达并表征了葡萄(Vitis vinifera)的功能性PME抑制剂VvPMEI1。VvPMEI1通常影响植物PMEs的活性,而对微生物PMEs无活性。通过表面等离子体共振研究的PMEI-PME相互作用动力学表明,该抑制剂在质外体pH值下与PME强烈相互作用,而随着pH值升高,复合物的稳定性降低。对VvPMEI1在不同葡萄组织和葡萄果实发育过程中的表达分析表明,这种抑制剂主要在浆果发育的早期阶段控制PME活性。在此阶段进行的蛋白质组学分析表明,一种PME同工型可能是VvPMEI1的作用靶点。