Role of apolipoprotein A-II in HDL structure-function
The experimental approachBy mutating single amino acids or deleting entire regions in apoA-II sequence we are searching for the most important structure/function determinants of the protein. We also label apoA-II at specific positions with fluorophores to investigate how the protein self-associates and associates with HDL. Experimentally we use mutagenesis techniques and our intein-based expression and purification system to produce specific apoA-II variants. The proteins are characterized structurally by circular dichroism and fluorescence spectroscopies, and mass-spectrometry. Interaction with other proteins on HDL is investigated by fluorescence spectroscopy. The biological activity of HLD reconstituted using apoA-II variants is also evaluated using cell culture techniques.