Structure and dynamics of acrylodan labeled αA-crystallin tetramer formed in the presence of a bile salt (sodium deoxycholate, NaDC) has been studied using fluorescence correlation spectroscopy (FCS) and femtosecond up-conversion techniques. Using FCS it is shown that, the diffusion constant (Dt) of the αA-crystallin oligomer (mass ∼800 kDa) increases from ∼35 μm2 s−1 to ∼68 μm2 s−1. This corresponds to a decrease in hydrodynamic radius (rh) from ∼6.9 nm to ∼3.3 nm. This corresponds to about 10-fold decrease in molecular mass to ∼80 kDa and suggests formation of a tetramer (since mass of αA-crystallin monomer is ∼20 kDa). The steady state emission maximum and average solvation time (〈τs〉) of acrylodan labeled at cysteine 131 position of αA-crystallin is markedly affected on addition of NaDC, while the tryptophan (trp-9) becomes more exposed. This suggests that NaDC binds near the cys-131 and makes the terminal region of αA-crystallin exposed. This may explain the enhanced auto-phosphorylation activity of αA-crystallin near the terminus of the 173 amino acid protein (e.g., at the threonine 13, serine 45, or serine 169 and 172) and suggests that phosphorylation at ser-122 (close to cys-131) is relatively less important.
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