The effect of dextran on subunit exchange of the molecular chaperone alphaA-crystallin.
Alpha-crystallin, a member of small heat shock protein (sHsp) family, is comprised of alphaA and alphaB subunits and acts as a molecular chaperone by interacting with unfolding proteins to prevent their aggregation. The alphaA-crystallin homopolymer consists of 30-40 subunits that are undergoing dynamic exchange. In vivo, alpha-crystallin elicits its chaperone action in a crowded cellular environment (e.g. in the lens). In vitro, inert molecular crowding agents (e.g. dextran) are often used to mimic crowded conditions. In this study, it was found that alpha-crystallin and alphaA-crystallin are poorer chaperones in the presence of dextran. Using fluorescence resonance energy transfer, it is shown that the alphaA-crystallin subunit exchange rate strongly increases with temperature. Binding of reduced ovotransferrin to alphaA-crystallin markedly decreases the rate of subunit exchange, as does the presence of dextran. In addition, in the presence of dextran the effect of reduced ovotransferrin on decreasing the rate of subunit exchange of alphaA-crystallin is greater than in the absence of dextran. Under the conditions of molecular crowding, the alphaA-crystallin subunit exchange rate is not temperature-dependent. In the absence of dextran, the exchange rate of alphaA-crystallin subunits correlates with its chaperone efficiency, i.e. the chaperone ability of alphaA-crystallin increases with temperature. However in the presence of dextran, the temperature dependence of the chaperone ability of alphaA-crystallin is eliminated.
Ghahghaei
Arezou
A
Department of Chemistry, University of Wollongong, Northfields Avenue, Wollongong, NSW 2522, Australia.
Rekas
Agata
A
Price
William E
WE
Carver
John A
JA
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D003207:Conalbumin / Q000494:pharmacology; D003911:Dextrans / Q000494:pharmacology*; D031541:Fluorescence Resonance Energy Transfer; D006358:Hot Temperature; D013050:Spectrometry, Fluorescence; D038202:alpha-Crystallin A Chain / Q000737:chemistry* / Q000187:drug effects
D003911:Dextrans; D038202:alpha-Crystallin A Chain; D003207:Conalbumin