Structure and expression of an inhibitor of fungal polygalacturonases from tomato.
A polygalacturonase inhibitor protein (PGIP) was characterized from tomato fruit. Differential glycosylation of a single polypeptide accounted for heterogeneity in concanavalin A binding and in molecular mass. Tomato PGIP had a native molecular mass of 35 to 41 kDa, a native isoelectric point of 9.0, and a chemically deglycosylated molecular mass of 34 kDa, suggesting shared structural similarities with pear fruit PGIP. When purified PGIPs from pear and tomato were compared, tomato PGIP was approximately twenty-fold less effective an inhibitor of polygalacturonase activity isolated from cultures of Botrytis cinerea. Based on partial amino acid sequence, polymerase chain reaction products and genomic clones were isolated and used to demonstrate the presence of PGIP mRNA in both immature and ripening fruit as well as cell suspension cultures. Nucleotide sequence analysis indicates that the gene, uninterrupted by introns, encodes a predicted 36.5 kDa polypeptide containing amino acid sequences determined from the purified protein and sharing 68% and 50% amino acid sequence identity with pear and bean PGIPs, respectively. Analysis of the PGIP sequences also revealed that they belong to a class of proteins which contain leucine-rich tandem repeats. Because these sequence domains have been associated with protein-protein interactions, it is possible that they contribute to the interaction between PGIP and fungal polygalacturonases.
Stotz
H U
HU
Department of Pomology, University of California, Davis 95616.
Contos
J J
JJ
Powell
A L
AL
Bennett
A B
AB
Labavitch
J M
JM
D000595:Amino Acid Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000378:metabolism; D005638:Fruit / Q000378:metabolism*; D003904:Mitosporic Fungi / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D010946:Plants, Medicinal; D011096:Polygalacturonase / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D011499:Protein Processing, Post-Translational; D012091:Repetitive Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid; D014675:Vegetables / Q000378:metabolism*
C081479:PGIP protein, plant; D010446:Peptide Fragments; D010940:Plant Proteins; D004247:DNA; D011096:Polygalacturonase