Recombinant wheat antifungal PR4 proteins expressed in Escherichia coli.
PR proteins are soluble and host-coded molecules with antifungal activity induced by a variety of agents. Wheat contains several PR proteins and among them are those of the class 4 coded wheatwin1 and wheatwin2; the two native proteins have been isolated from wheat kernel and the coding cDNA clones have been recently characterized. Herein, we report the expression of recombinant wheatwin1 and wheatwin2 in Escherichia coli-insoluble fractions; a new protocol for the purification in high yields and correct processing of the two proteins was developed. The recombinant proteins have molecular weights identical to that of the native proteins, indicating that the removal of the N-terminal methionine and cyclization of glutamine to pyroglutamate was complete. Both recombinant proteins inhibited in vitro the growth of Fusarium culmorum exhibiting antifungal properties similar to those of the native proteins.
Caruso
C
C
Dipartimento di Agrobiologia e Agrochimica, Università della Tusica, Italy. caruso@unitus.it
Bertini
L
L
Tucci
M
M
Caporale
C
C
Nobile
M
M
Leonardi
L
L
Buonocore
V
V
D000935:Antifungal Agents / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics*; D005670:Fusarium / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure; D002479:Inclusion Bodies / Q000378:metabolism; D018613:Microscopy, Confocal; D008958:Models, Molecular; D008970:Molecular Weight; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014170:Transformation, Genetic; D014908:Triticum
D000935:Antifungal Agents; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C062880:Win-1 proteins, plant; C053376:pathogenesis-related proteins, plant