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A single amino acid mutation affects elicitor and expansins-like activities of cerato-platanin, a non-catalytic fungal protein
- Source :
- PLoS ONE, PLoS ONE, Vol 12, Iss 5, p e0178337 (2017)
- Publication Year :
- 2016
-
Abstract
- 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/β 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.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Fungal Structure
Mutant
lcsh:Medicine
Plant Science
medicine.disease_cause
01 natural sciences
Biochemistry
chemistry.chemical_compound
Filter Paper
Urea
lcsh:Science
Plant Proteins
Fungal protein
Multidisciplinary
Organic Compounds
Cerato-platanin
Biological activity
Plants
Recombinant Proteins
Elicitor
Laboratory Equipment
Chemistry
Experimental Organism Systems
Plant Physiology
Host-Pathogen Interactions
Physical Sciences
Engineering and Technology
Research Article
Arabidopsis Thaliana
Equipment
Brassica
Mycology
Biology
Research and Analysis Methods
Fungal Proteins
03 medical and health sciences
Expansin
Model Organisms
Plant and Algal Models
Phytoalexins
medicine
Escherichia coli
Plant Defenses
Cellulose
lcsh:R
Pathogen-Associated Molecular Pattern Molecules
Organic Chemistry
Chemical Compounds
Organisms
Fungi
Biology and Life Sciences
Proteins
Plant Disease Resistance
Plant Pathology
030104 developmental biology
chemistry
Amino Acid Substitution
cerato-platanin
defence elicitor, E. coli SHuffle, expansin-like protein, PAMP, ROS
lcsh:Q
010606 plant biology & botany
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 12
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....f932665c775e6d12c207fb774c5952a4