Insulin amyloid fibrillation at above 100 degrees C: new insights into protein folding under extreme temperatures.
To investigate the folding behavior of amyloidogenic proteins under extreme temperatures, the kinetics of fibrillation and accompanying secondary structure transitions of bovine insulin were studied for temperatures ranging up to 140 degrees C. The presence of extreme heat stress had traditionally been associated with irreversible denaturation of protein while the initial steps of such a denaturation process may be common with a fibril formation pathway of amyloidogenic proteins. The present work demonstrates the ability of insulin to form amyloid fibrils at above 100 degrees C. Amyloid formation was gradually replaced by random coil generation after approximately 80 degrees C until no amyloid was detected at 140 degrees C. The morphology of insulin amyloid fibrils underwent sharp changes with increasing the temperature. The dependence of amyloid formation rate on incubation temperature followed non-Arrhenius kinetics, which is explained by temperature-dependent enthalpy change for amyloid formation. The intermediate stage of amyloid formation and random coil generation consisted of a partially folded intermediate common to both pathways. The fully unfolded monomers in random coil conformation showed partial reversibility through this intermediate by reverting back to the amyloid pathway when formed at 140 degrees C and incubated at 100 degrees C. This study highlights the non-Arrhenius kinetics of amyloid fibrillation under extreme temperatures, and elucidates its intermediate stage common with random coil formation.
Department of Chemical and Materials Engineering, Arizona State University, Tempe, AZ 85287, USA.
Anubhav
A
Arora
Chanki
C
Ha
Chan Beum
CB
Park
D000682:Amyloid / Q000737:chemistry*; D002942:Circular Dichroism; D007328:Insulin / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013696:Temperature
D000682:Amyloid; D007328:Insulin