title: Monitoring protein aggregation during thermal unfolding in circular dichroism experiments.
abstract: Thermal unfolding monitored by spectroscopy or calorimetry is widely used to determine protein stability. Equilibrium thermodynamic analysis of such unfolding is often hampered by its irreversibility, which usually results from aggregation of thermally denatured protein. In addition, heat-induced protein misfolding and aggregation often lead to formation of amyloid-like structures. We propose a convenient method to monitor in real time protein aggregation during thermal folding/ unfolding transition by recording turbidity or 90 degrees light scattering data in circular dichroism (CD) spectroscopic experiments. Since the measurements of turbidity and 90 degrees light scattering can be done simultaneously with far- or near-UV CD data collection, they require no additional time or sample and can be directly correlated with the protein conformational changes monitored by CD. The results can provide useful insights into the origins of irreversible conformational changes and test the linkage between protein unfolding or misfolding and aggregation in various macromolecular systems, including globular proteins and protein-lipid complexes described in this study, as well as a wide range of amyloid-forming proteins and peptides.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, W329, 715 Albany Street, Boston, MA 02118, USA.
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Benjwal
authors.forename: Shikha
authors.initials: S
authors.lastname: Verma
authors.forename: Klaus-Heinrich
authors.initials: KH
authors.lastname: Röhm
authors.forename: Olga
authors.initials: O
authors.lastname: Gursky
mesh_terms: D001215:Asparaginase / Q000737:chemistry; D002942:Circular Dichroism / Q000379:methods*; D008027:Light; D008074:Lipoproteins / Q000737:chemistry; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
chemical_list: D008074:Lipoproteins; D011506:Proteins; D001215:Asparaginase