Publication Details

Morris, A. M. & Aquilina, J. Andrew. (2010). Evidence for specific subunit distribution and interactions in the quaternary structure of α-crystallin. Proteins: Structure, Function, and Bioinformatics, 78 (11), 2546-2553.


The quaternary structure of α-crystallin is dynamic, a property which has thwarted crystallographic efforts towards structural characterization. In this study, we have used collision-induced dissociation mass spectrometry to examine the architecture of the polydisperse assemblies of α-crystallin. For total α-crystallin isolated directly from fetal calf lens using size-based chromatography, the αB-crystallin subunit was found to be preferentially dissociated from the oligomers, despite being significantly less abundant overall than the αA-crystallin subunits. Furthermore, upon mixing molar equivalents of purified αA- and αB-crystallin, the levels of their dissociation were found to decrease and increase, respectively, with time. Interestingly though, dissociation of subunits from the αA- and αB-crystallin homo-oligomers was comparable, indicating that strength of the αA:αA, and αB:αB subunit interactions are similar. Taken together, these data suggest that differences in the number of subunit contacts in the mixed assemblies give rise to the disproportionate dissociation of αB-crystallin subunits. Limited proteolysis mass spectrometry was also used to examine changes in protease accessibility during subunit exchange. The C-terminus of αA-crystallin was more susceptible to proteolytic attack in homo-oligomers than that of αB-crystallin. As subunit exchange proceeded, proteolysis of the αA-crystallin C-terminus increased, indicating that in the hetero-oligomeric form this tertiary motif is more exposed to solvent. These data were used to propose a refined arrangement for the interactions of the α-crystallin domains and C-terminal extensions of subunits within the α-crystallin assembly. In particular we propose that the palindromic IPI motif of αB-crystallin gives rise to two orientations of the C-terminus.


060101 Analytical Biochemistry



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