ABSTRACT
Antimutagenic activity on human RD cells was studied for beta-purothionin Tk-AMP-BP isolated from seeds of wheat Triticum kiharae, which has a higher stress resistance. Cadmium chloride at 5 x 10(-6) M was used as a mutagen. The numbers of DNA breaks in mutagen-treated and intact cells were inferred from the single-stranded to double-stranded DNA ratio and expressed as protection coefficients. The protective effect was simultaneously assayed for aqueous plant extracts known to possess antioxidant properties. Wheat thionin was the most active among all of the antimutagens examined; its protection coefficient reached 85-88% at micromolar peptide concentrations (8-32 microg/ml). Thus, wheat beta-purothionin was for the first time demonstrated to be highly efficient in protecting human cell DNA from the damaging effect of cadmium chloride.
Subject(s)
Antimicrobial Cationic Peptides/pharmacology , Antimutagenic Agents/pharmacology , DNA Breaks/drug effects , Plant Proteins/pharmacology , Seeds/chemistry , Triticum/chemistry , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/isolation & purification , Antimutagenic Agents/chemistry , Antimutagenic Agents/isolation & purification , Cadmium Chloride/pharmacology , Cell Line , Dose-Response Relationship, Drug , Humans , Mutagens/pharmacology , Plant Proteins/chemistry , Plant Proteins/isolation & purificationABSTRACT
A novel antifungal peptide, LAMP-Ia, was isolated from sand-elymus (Leymus arenarius) seeds. Expression of a synthetic gene encoding this peptide in Escherichia coli cells was obtained. The target peptide was expressed as a fusion with thioredoxin. Identity of the recombinant peptide to native LAMP-Ia was confirmed by chromatography, mass spectrometry, and amino acid sequencing. LAMP-Ia displayed a high inhibitory activity in respect of a number of phytopathogenic fungi in in vitro assays, which opens up possibilities for the gene encoding it to be used for genetic transformation of plants and for engineering pathogen-resistant crops.