Structural requirements of endopolygalacturonase for the interaction with PGIP (polygalacturonase-inhibiting protein).
To invade a plant tissue, phytopathogenic fungi produce several cell wall-degrading enzymes; among them, endopolygalacturonase (PG) catalyzes the fragmentation and solubilization of homogalacturonan. Polygalacturonase-inhibiting proteins (PGIPs), found in the cell wall of many plants, counteract fungal PGs by forming specific complexes with them. We report the crystal structure at 1.73 A resolution of PG from the phytopathogenic fungus Fusarium moniliforme (FmPG). The structure of FmPG was useful to study the mode of interaction of the enzyme with PGIP-2 from Phaseolus vulgaris. Several amino acids of FmPG were mutated, and their contribution to the formation of the complex with PGIP-2 was investigated by surface plasmon resonance. The residues Lys-269 and Arg-267, located inside the active site cleft, and His-188, at the edge of the active site cleft, are critical for the formation of the complex, which is consistent with the observed competitive inhibition of the enzyme played by PGIP-2. The replacement of His-188 with a proline or the insertion of a tryptophan after position 270, variations that both occur in plant PGs, interferes with the formation of the complex. We suggest that these variations are important structural requirements of plant PGs to prevent PGIP binding.
Federici
L
L
Dipartimento di Scienze Biochimiche, Università di Roma La Sapienza, Piazzale le Aldo Moro 5, 00185 Rome, Italy.
Caprari
C
C
Mattei
B
B
Savino
C
C
Di Matteo
A
A
De Lorenzo
G
G
Cervone
F
F
Tsernoglou
D
D
D001483:Base Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D017931:DNA Primers; D005670:Fusarium / Q000201:enzymology; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010940:Plant Proteins / Q000378:metabolism*; D011096:Polygalacturonase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D020349:Surface Plasmon Resonance
D017931:DNA Primers; C081479:PGIP protein, plant; D010940:Plant Proteins; D011096:Polygalacturonase