RESUMO
Grapevine (Vitis vinifera L.) is a crop of major economic importance. However, grapevine yield is guaranteed by the massive use of pesticides to counteract pathogen infections. Under temperate-humid climate conditions, downy mildew is a primary threat for viticulture. Downy mildew is caused by the biotrophic oomycete Plasmopara viticola Berl. & de Toni, which can attack grapevine green tissues. In lack of treatments and with favourable weather conditions, downy mildew can devastate up to 75% of grape cultivation in one season and weaken newly born shoots, causing serious economic losses. Nevertheless, the repeated and massive use of some fungicides can lead to environmental pollution, negative impact on non-targeted organisms, development of resistance, residual toxicity and can foster human health concerns. In this manuscript, we provide an innovative approach to obtain specific pathogen protection for plants. By using the yeast two-hybrid approach and the P. viticola cellulose synthase 2 (PvCesA2), as target enzyme, we screened a combinatorial 8 amino acid peptide library with the aim to identify interacting peptides, potentially able to inhibit PvCesa2. Here, we demonstrate that the NoPv1 peptide aptamer prevents P. viticola germ tube formation and grapevine leaf infection without affecting the growth of non-target organisms and without being toxic for human cells. Furthermore, NoPv1 is also able to counteract Phytophthora infestans growth, the causal agent of late blight in potato and tomato, possibly as a consequence of the high amino acid sequence similarity between P. viticola and P. infestans cellulose synthase enzymes.
Assuntos
Aptâmeros de Peptídeos/farmacologia , Glucosiltransferases/antagonistas & inibidores , Oomicetos/efeitos dos fármacos , Doenças das Plantas/terapia , Proteínas de Plantas/antagonistas & inibidores , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Sequência de Aminoácidos , Celulose/biossíntese , Glucosiltransferases/química , Oomicetos/enzimologia , Oomicetos/ultraestrutura , Biblioteca de Peptídeos , Fotossíntese , Phytophthora infestans/efeitos dos fármacos , Phytophthora infestans/enzimologia , Phytophthora infestans/ultraestrutura , Doenças das Plantas/parasitologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Proteínas de Plantas/química , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Solanum tuberosum , Técnicas do Sistema de Duplo-Híbrido , VitisRESUMO
The theoretical basis behind the ability of constrained Cα-tetrasubstituted amino acids (CTAAs) to induce stable helical conformations has been studied through Replica Exchange Molecular Dynamics Potential of Mean Force Quantum Theory of Atoms In Molecules calculations on Ac-l-Ala-CTAA-l-Ala-Aib-l-Ala-NHMe peptide models. We found that the origin of helix stabilization by CTAAs can be ascribed to at least two complementary mechanisms limiting the backbone conformational freedom: steric hindrance predominantly in the (+x,+y,-z) sector of a right-handed 3D Cartesian space, where the z axis coincides with the helical axis and the Cα of the CTAA lies on the +y axis (0,+y,0), and the establishment of additional and relatively strong C-H···O interactions involving the CTAA.
Assuntos
Aminoácidos/química , Estrutura Secundária de Proteína , Ligação de Hidrogênio , Estabilidade ProteicaRESUMO
Enantiopure constrained 1-aminocyclopentane-1,2,4-tricarboxylic acids containing the glutamic acid skeleton were prepared as two diastereomers characterized by having the carboxylic groups in position two and four cis-oriented to each other and trans with respect to 1-carboxylic group and all cis-oriented carboxylic groups, respectively. A biochemical screening of activity of the above amino acids was investigated on glutamate and 5-HT receptors to find a possible metabotropic agonist, acting on the serotoninergic system.