Amyloidosis Proteins and Fibril Formation Studies

We perform biochemical and biophysical studies on AL amyloidosis proteins predominantly from recombinant sources (E. coli) as well as urine- and cell-line-derived proteins. Initial characterization of protein identity, purity and oligomerization state is done using mass spectrometry, amino acid analysis, size exclusion chromatography and, in some cases, analytical ultracentrifugation. We study protein structure thermodynamic stability by thermal and chemical denaturation using circular dichroism and fluorescence spectroscopy. We study amyloid formation in using vitro thioflavine T fluorescence as well as electron microscopy.

We are interested in the role of accessory molecules and cytotoxicity in protein structure, stability, and amyloid formation kinetics in vivo and in vitro. These accessory factors include extracellular matrix components such as glycosaminoglycans as well as lipids, lipoproteins and other serum components.

We are currently studying the possible toxic effect of AL amyloidosis protein oligomeric species and amyloid fibrils formed in the presence of different external factors in cardiomyocytes. We are also studying light chain internalization into cardiomyocytes by fluorescently tagging our AL amyloidosis proteins and following their uptake rates and intracellular localization via confocal microscopy.