Doctor of Philosophy
Department of Chemistry
Cooper, Philip G., 1H NMR spectroscopic studies on bovine lens crystallins, Doctor of Philosophy thesis, Department of Chemistry, University of Wollongong, 1993. http://ro.uow.edu.au/theses/1105
The research in this thesis involved the separation and isolation of crystallin proteins from bovine lenses and the analysis of the protein isolates by 1 H two dimensional NMR spectroscopy. Sequential assignments were made for extended and flexible terminal regions of βB2-, γs- and γB-crystallin, which assume random coil conformations. NMR spectral analysis showed that different conformations of the terminal extensions and flexible regions within the domain regions exist between the p subunits. This is consistent with separate roles for each subunit in aggregate formation. It is shown that the C-terminal extension of the βB2 subunit is involved in total β aggregation at in vivo concentrations. Evidence is also presented to show that the γs-crystallin has conformational attributes intermediate to those of the β- and γ-crystallins. NMR spectra acquired for a range of crystallin homogenates give supporting evidence for supra-molecular interactions between the crystallins with specific interactions deduced for the assigned terminal extensions. Based on these results a model is proposed of how the crystallins maintain short order interactions consistent with lens transparency.