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Expression, purification, crystallization and X-ray diffraction studies of a novel root-induced secreted protein from Trichoderma virens
- Source :
- Acta Crystallogr F Struct Biol Commun
- Publication Year :
- 2020
- Publisher :
- International Union of Crystallography (IUCr), 2020.
-
Abstract
- Small secreted cysteine-rich proteins (SSCPs) from fungi play an important role in fungi–host interactions. The plant-beneficial fungi Trichoderma spp. are in use worldwide as biocontrol agents and protect the host plant from soil-borne as well as foliar pathogens. Recently, a novel SSCP, Tsp1, has been identified in the secreted protein pool of T. virens and is overinduced upon its interaction with the roots of the maize plant. The protein was observed to be well conserved in the Ascomycota division of fungi, and its homologs are present in many plant-pathogenic fungi such as Fusarium oxysporum and Magnaporthe oryzae. However, none of these homologs have yet been characterized. Recombinant Tsp1 protein has been expressed and purified using an Escherichia coli expression system. The protein, with four conserved cysteines, forms a dimer in solution as observed by size-exclusion chromatography. The dimerization, however, does not involve disulfide bonds. Circular-dichroism data suggested that the protein has a β-strand-rich secondary structure that matched well with the secondary structure predicted using bioinformatics methods. The protein was crystallized using sodium malonate as a precipitant. The crystals diffracted X-rays to 1.7 Å resolution and belonged to the orthorhombic space group P212121 (R meas = 5.4%), with unit-cell parameters a = 46.3, b = 67.0, c = 173.2 Å. The Matthews coefficient (V M) of the crystal is 2.32 Å3 Da−1, which corresponds to nearly 47% solvent content with four subunits of Tsp1 protein in the asymmetric unit. This is the first report of the structural study of any homolog of the novel Tsp1 protein. These structural studies will help in understanding the classification and function of the protein.
- Subjects :
- Models, Molecular
0106 biological sciences
Protein Conformation
Hypocrea
Dimer
Biophysics
Sequence Homology
Crystallography, X-Ray
medicine.disease_cause
01 natural sciences
Biochemistry
Research Communications
law.invention
Fungal Proteins
03 medical and health sciences
chemistry.chemical_compound
Structural Biology
law
Fusarium oxysporum
Genetics
medicine
Amino Acid Sequence
Cysteine
Escherichia coli
Protein secondary structure
030304 developmental biology
0303 health sciences
biology
Ascomycota
Chemistry
food and beverages
Condensed Matter Physics
biology.organism_classification
Recombinant Proteins
Malonate
Recombinant DNA
Function (biology)
010606 plant biology & botany
Subjects
Details
- ISSN :
- 2053230X
- Volume :
- 76
- Database :
- OpenAIRE
- Journal :
- Acta Crystallographica Section F Structural Biology Communications
- Accession number :
- edsair.doi.dedup.....49fe184f74c553b9343ae1bdf994bd51