The study of the properties and mechanisms of chaperone functioning is not only fundamental, but also of practical interest in the development of drugs for the prevention and treatment of neurodegenerative diseases. In this study, using the dynamic light scattering the inhibitory effect of Skp chaperone on the process of assembling the recombinant OmpF porin from the outer membrane of Yersinia pseudotuberculosis (rOmpF) was demonstrated. The distribution curves of the protein particles and the mean value of their hydrodynamic radii (RH) were obtained from the denatured state (in 8 M urea, RH 8,2 nm) into the aqueous solution at acidic and alkaline pH values. It was shown that at pH 5,0, under conditions close to the isoelectric point of the protein, rOmpF quickly and irreversibly aggregated. Thus, with a 10-fold dilution of a denatured rOmpF solution with a pH 5,0 buffer, formation of particles with average RH 53 nm was already observed in the first minutes, after an hour they increased to the size of sedimenting particles (1076 nm). In the presence of rSkp under these conditions, the particle size rOmpF (average RH 50,6 nm) only slightly changed during the day (average RH of 61,7 nm). At pH 8,0, reversible self-association of rOmpF was observed with the formation of a wide size distribution of the protein particles (from 8 to 80-60 nm), which changed insignificantly with time. Under these conditions, Skp formed a sufficiently stable complex with rOmpF, the particle size of which (mean RH 5,1 nm) was preserved during the day.
chaperone Skp, recombinant porin OmpF, Yersinia pseudotuberculosis, outer membrane protein, aggregation of proteins, protein-protein interactions, dynamic light scattering
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