ABSTRACT
Crotamine is one of the main constituents of the venom of the South American rattlesnake Crotalus durissus terrificus. A common gene ancestry and structural similarity with the antimicrobial ß-defensins (identical disulfide bond pattern and highly positive net charge) suggested potential antimicrobial activities for this snake toxin. Although crotamine demonstrated low activity against both Gram-positive and Gram-negative bacteria, a pronounced antifungal activity was observed against Candida spp., Trichosporon spp., and Cryptococcus neoformans. Crotamine's selective antimicrobial properties, with no observable hemolytic activity, stimulated us to evaluate the potential applications of this polypeptide as an antiyeast or candicidal agent for medical and industrial application. Aiming to understand the mechanism(s) of action underlying crotamine antimicrobial activity and its selectivity for fungi, we present herein studies using membrane model systems (i.e., large unilamellar vesicles, LUVs, and giant unilamellar vesicles, GUVs), with different phospholipid compositions. We show here that crotamine presents a higher lytic activity on negatively charged membranes compared with neutral membranes, with or without cholesterol or ergosterol content. The vesicle burst was not preceded by membrane permeabilization as is generally observed for pore forming peptides. Although such a property of disrupting lipid membranes is very important to combat multiresistant fungi, no inhibitory activity was observed for crotamine against biofilms formed by several Candida spp. strains, except for a limited effect against C. krusei biofilm.
Subject(s)
Crotalid Venoms/chemistry , Crotalus/metabolism , Unilamellar Liposomes/chemistry , Amino Acid Sequence , Animals , Antifungal Agents/pharmacology , Crotalid Venoms/metabolism , Crotalid Venoms/toxicity , Fungi/drug effects , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Microscopy , Molecular Sequence Data , Unilamellar Liposomes/metabolismABSTRACT
Water at room temperature is not simply a medium for which uniform properties can always be assumed. Water close to solid hydrophobic or hydrophilic surfaces has elasticity, which is measured by monitoring the quartz crystal microbalance (QCM) resonant frequency and resistance. Small additions of salt are shown to modify this elasticity. Furthermore, near the hydrophobic QCM gold electrode, undersaturated aqueous NaCl solutions present a high concentration of ion pairs, which is confirmed by atomic force microscopy through force versus distance measurements.
