Structure, supramolecular association and interaction of patho-active presenilin and betaAPP domains by high resolution mass spectrometry

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In this project the detailed structures, interactions and supramolecular association of b-amyloid precursor protein (bAPP) and presinilin(s) (PS), two key molecules for Alzheimer's disease, will be evaluated using new methods of protein mass spectrometry (electrospray / ESI; matrix-assisted laser desorption / MALDI). In particular, Fourier-transform-ICR-MS will be used as a major bioanalytical tool providing ultrahigh sensitivity and resolution for structural studies. bAPP polypeptides spanning the Ab transmembrane sequences, and PS transmembrane spanning regions 6 and 7 will be prepared by chemical synthesis to include sequence mutations at the presumed proteolytic cleavage site. Coupling of MS with the mass spectrometric epitope mapping developed in our laboratory will be employed to study interactions of PS with Ab sequences. A new affinity proteome-MS approach using micro-capillary- and chip-immobilised antibodies will be applied to identify intracellular association products and components of the PS complex. Furthermore, the synthesis of large Ab-containing polypeptides should provide suitable models for future studies of pathophysiological interactions of PS and bAPP.