[PMID]: | 28542638 |
[Au] Autor: | Luti S; Martellini F; Bemporad F; Mazzoli L; Paoli P; Pazzagli L |
[Ad] Endereço: | Department of Biomedical, Experimental and Clinical Sciences, University of Florence, Florence, Italy. |
[Ti] Título: | A single amino acid mutation affects elicitor and expansins-like activities of cerato-platanin, a non-catalytic fungal protein. |
[So] Source: | PLoS One;12(5):e0178337, 2017. |
[Is] ISSN: | 1932-6203 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | Cerato-platanin (CP) is a non-catalytic, cysteine-rich protein, the first member of the cerato-platanin family. It is a single-domain protein with a double Ψ/ß barrel domain resembling the D1 domain of plant and bacterial expansins. Similarly to expansins, CP shows a cell wall-loosening activity on cellulose and can be defined as an expanisin-like protein, in spite of the missing D2 domain, normally present in plant expansins. The weakening activity shown on cellulose may facilitate the CP-host interaction, corroborating the role of CP in eliciting plant defence response. Indeed, CP is an elicitor of primary defences acting as a Pathogen-Associated Molecular Patterns (PAMP). So far, structure-function relationship study has been mainly performed on the bacterial BsEXLX1 expansin, probably due to difficulties in expressing plant expansins in heterologous systems. Here, we report a subcloning and purification method of CP in the engineered E. coli SHuffle cells, which proved to be suitable to obtain the properly folded and biologically active protein. The method also enabled the production of the mutant D77A, rationally designed to be inactive. The wild-type and the mutated CP were characterized for cellulose weakening activity and for PAMP activity (i.e. induction of Reactive Oxygen Species synthesis and phytoalexins production). Our analysis reveals that the carboxyl group of D77 is crucial for expansin-like and PAMP activities, thus permitting to establish a correlation between the ability to weaken cellulose and the capacity to induce defence responses in plants. Our results enable the structural and functional characterization of a mono-domain eukaryotic expansin and identify the essential role of a specific aspartic residue in cellulose weakening. |
[Mh] Termos MeSH primário: |
Proteínas Fúngicas/genética
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[Mh] Termos MeSH secundário: |
Substituição de Aminoácidos Celulose/metabolismo Escherichia coli/genética Proteínas Fúngicas/fisiologia Interações Hospedeiro-Patógeno/genética Padrões Moleculares Associados a Patógenos/metabolismo Proteínas de Plantas/genética Proteínas de Plantas/fisiologia
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[Pt] Tipo de publicação: | JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Cerato-platanin protein, Ceratocystis fimbriata); 0 (Fungal Proteins); 0 (Pathogen-Associated Molecular Pattern Molecules); 0 (Plant Proteins); 0 (expansin protein, plant); 9004-34-6 (Cellulose) |
[Em] Mês de entrada: | 1709 |
[Cu] Atualização por classe: | 170919 |
[Lr] Data última revisão:
| 170919 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 170526 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1371/journal.pone.0178337 |
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