科研之友
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摘要
The effect of crowding on the chaperone-like activity of alpha-crystallin has been studied using aggregation of UV-irradiated glycogen phosphorylase b (Phb) from rabbit skeletal muscle as an aggregation test system. The merit of this test system is the possibility of testing agents that directly affect the stage of aggregation of the protein molecules. It was shown that the solution of Phb denatured by UV contained aggregates with a hydrodynamic radius of 10.4 nm. These aggregates are relatively stable at 20 degrees C; however, they reveal a tendency to stick further in the presence of crowding agents. The study of the effect of alpha-crystallin on the aggregation of UV-irradiated Phb in the presence of the crowding agents by dynamic light scattering at 37 degrees C showed that under crowding conditions the antiaggregation ability of alpha-crystallin was weakened. On the basis of the analytical ultracentrifugation, size-exclusion chromatography, and sodium dodecyl sulfate polyacrylamide gel electrophoresis data, the scheme of interaction of UV-irradiated Phb and alpha-crystallin has been proposed. It is assumed that chaperone target protein complexes of two types are formed, namely, the complexes of dissociated forms of alpha-crystallin with a protein substrate and high-mass alpha-crystallin-denatured protein complexes. The complexes of the first type reveal a weak propensity to aggregate even under crowding conditions. The complexes of the second type are characterized by the lower rate of aggregation in comparison with that of original UV-irradiated Phb. However, crowding stimulates the rate of aggregation of these complexes, resulting in the above-mentioned decrease in the chaperone-like activity of alpha-crystallin.
- 出版日期2011-12-13
