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Characterization of Two Novel Type I Ribosome-Inactivating
Proteins from the Storage Roots of the Andean Crop
Mirabilis
expansa1
Jorge M. Vivanco,
Brett J. Savary, and
Hector E. Flores*
Department of Plant Pathology and Biotechnology Institute, The
Pennsylvania State University, University Park, Pennsylvania 16802 (J.M.V., H.E.F.); and United States Department of
Agriculture-Agricultural Research Service, Eastern Region Research
Center, 600 East Mermaid Lane, Wyndmoor, Pennsylvania 19038 (B.J.S.)
Two novel type I
ribosome-inactivating proteins (RIPs) were found in the storage roots
of Mirabilis expansa, an underutilized Andean root crop.
The two RIPs, named ME1 and ME2, were purified to homogeneity by
ammonium sulfate precipitation, cation-exchange perfusion
chromatography, and C4 reverse-phase chromatography. The
two proteins were found to be similar in size (27 and 27.5 kD) by
sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their
isoelectric points were determined to be greater than pH 10.0. Amino
acid N-terminal sequencing revealed that both ME1 and ME2 had conserved
residues characteristic of RIPs. Amino acid composition and
western-blot analysis further suggested a structural similarity between
ME1 and ME2. ME2 showed high similarity to the Mirabilis
jalapa antiviral protein, a type I RIP. Depurination of yeast
26S rRNA by ME1 and ME2 demonstrated their ribosome-inactivating activity. Because these two proteins were isolated from roots, their
antimicrobial activity was tested against root-rot microorganisms, among others. ME1 and ME2 were active against several fungi, including Pythium irregulare, Fusarium oxysporum
solani, Alternaria solani, Trichoderma reesei, and Trichoderma
harzianum, and an additive antifungal effect of ME1 and ME2 was
observed. Antibacterial activity of both ME1 and ME2 was observed
against Pseudomonas syringae, Agrobacterium
tumefaciens, Agrobacterium radiobacter, and
others.
1
This work was supported by the McKnight
Foundation.
*
Corresponding author; e-mail hef1{at}psu.edu; fax 1-814-863-7217.
Plant Physiol. (1999) 119: 1447-1456
Copyright Clearance Center: 0032-0889/99/119//10
© 1999 American Society of Plant Physiologists
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