350 rub
Journal Technologies of Living Systems №8 for 2010 г.
Article in number:
Advantages of application of a pulsed XeCl laser (308 nm) for elucidation of aggregation
Keywords: eye lens crystallins photoaggregation effect of XeCl laser irradiation
Authors:
L.V. Soustov, E.V. Chelnokov, N.V. Sapogova, N.M. Bityurin, M.A.Ostrovsky
Abstract:
Different denaturant factors, including UV radiation, can cause crystallin aggregation that leads to increase of lens opacity promoting cataract development. Application of pulsed UV laser radiation of a XeCl laser within the wide range of pulse energy densities w and pulse repetition rates F can be regarded as an effective methodical tool when studying the protein photo-aggregation. In known to us papers the elucidation of the photoaggregation was carried out either by means of the continuous UV irradiation of a mercury lamp with 10 nm bandwidth or by using the pulsed laser radiation at the fixed pulse energy density w and pulse repetition rate F. As it is shown in the present paper the pulsed monochromatic laser radiation at wavelength 308 nm is more effective for protein aggregation than the mercury lamp radiation. Being narrowed down to 10 nm bandwidth mercury lamp radiation has significantly less power than laser radiation thus providing lower protein aggregation rate and requiring more long-run experiment. The paper presents the results of the study on the laser-induced photoaggregation of water-soluble alpha-, beta-, gamma-crystallins, and carbonic anhydrase. The former are the main eye lens proteins. The irradiation was performed by a XeCl laser at 308 nm within the wide range of pulse energy densities w and pulse repetition rates F. The nonlinear dependencies of aggregation efficiency on the values of w, F and the concentration of protein solution are observed. The theo-retical model that qualitatively describes the experimental results is developed. It should be noted that the devised method is also applicable for photoaggegation research of any water-soluble proteins. Besides, such model experiments are promising for searching for new compositions reducing the protein aggregation. Chaperon-like activity of α-crystallin in βL-crystallin aggregation process is studied. The in-fluence of some short-chain peptides on aggregation efficiency of crystallins irradiated by a pulsed XeCl laser is tested. The result of in vitro elucidation of a new peptide composition is presented. It is shown that developed composition provides greater delay of crystallin photoaggregation than known anticata-ract preparations. The developed composition of peptides has been investigated in vivo with expe-rimental animals. An increase of the delay time of UV-induced cataract development provided by the composition in the rat is observed. The research was carried out to study the dependence of the rate of UV induced aggregation of alpha- and beta-crystallin mixture on the developed composition ingredient proportion. A new phenomenon of «postaggregation» is discovered when following the aggregation of car-bonic anhydrase after cutting off the laser irradiation of the solution of the protein. Similar expe-riments with alpha-, beta- and gamma-crystallins are under progress.
Pages: 29-37
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