Cloned (Comment) | Organism |
---|---|
cloning in Escherichia coli strain TOP10F, expression in Pichia pastoris strain X-33 and secretion to the culture medium | Fusarium verticillioides |
recombinant enzyme expression | Fusarium phyllophilum |
Protein Variants | Comment | Organism |
---|---|---|
A274T | site-directed mutagenesis, the mutation causes a marked loss of inhibition by PvPGIP2 of 150fold | Fusarium phyllophilum |
K116E | site-directed mutagenesis, the mutant shows no inhibition by PvPGIP2 | Fusarium phyllophilum |
K310T | site-directed mutagenesis | Fusarium phyllophilum |
L303E | site-directed mutagenesis | Fusarium verticillioides |
L303E | site-directed mutagenesis, the mutant enzyme is inhibited by Inhibitor mutant PvPGIP2.Q224K in contrast to the wild-type enzyme | Fusarium phyllophilum |
N121K | site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 | Fusarium phyllophilum |
Q124P | site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 | Fusarium phyllophilum |
S120N | site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 | Fusarium phyllophilum |
S120N/N121K/S122D | site-directed mutagenesis, the triple mutant is inhibited with a 25fold reduced efficiency by PvPGIP2 | Fusarium phyllophilum |
S122D, | site-directed mutagenesis, the mutation only slightly affects inhibition by PvPGIP2 | Fusarium phyllophilum |
S363K | site-directed mutagenesis | Fusarium phyllophilum |
T274A | site-directed mutagenesis | Fusarium verticillioides |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | inability of the enzyme from Fusarium phyllophilum to be inhibited by polygalacturonase-inhibiting protein 2, PGIP2, from Phaseolus vulgaris pv. Pinto and recombinatly expressed in Pichia pastoris X33, or any other polygalacturonase-inhibiting protein, small angle X-ray scattering interaction analysis of structural basis, overview | Fusarium verticillioides | |
polygalacturonase-inhibiting protein 2 | PGIP2, from Phaseolus vulgaris pv. Pinto, recombinantly expressed in Pichia pastoris X33, forms a complex with the enzyme from Colletotrichum lupini hindering substrate binding in the enzyme active site cleft, small angle X-ray scattering interaction analysis, and inhibition kinetics, overview | Colletotrichum lupini | |
polygalacturonase-inhibiting protein 2 | PGIP2, from Phaseolus vulgaris pv. Pinto, recombinantly expressed in Pichia pastoris X33, forms a complex with the enzyme from Fusarium phyllophilum hindering substrate binding in the enzyme active site cleft, residue A274 plays a key role in the interaction, small angle X-ray scattering interaction analysis, and inhibition kinetics, overview. Residue Q224 of the inhibitor is crucial for the interaction with the enzyme. Inhibitor mutant PvPGIP2.Q224K variant is unable to inhibit the enzyme, while inhibitor mutant PvPGIP2.Q224K retains unaltered inhibitory capability towards the enzyme | Fusarium phyllophilum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Colletotrichum lupini | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Colletotrichum lupini | - |
- |
- |
Colletotrichum lupini SHK78813 | - |
- |
- |
Fusarium phyllophilum | - |
- |
- |
Fusarium verticillioides | - |
- |
- |
Fusarium verticillioides 62264 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
native extracellular enzyme from culture medium | Colletotrichum lupini |
recombinant enzyme | Fusarium phyllophilum |
recombinant extracellular enzyme from Pichia pastoris strain X-33 culture medium by ultrafiltration and dialysis, anion and cation exchange chromatography, dialysis, and hydrophobic interaction chromatography | Fusarium verticillioides |
Synonyms | Comment | Organism |
---|---|---|
CluPG1 | - |
Colletotrichum lupini |
FgPG | - |
Fusarium verticillioides |
FpPG | - |
Fusarium phyllophilum |