Off-pathway aggregation can inhibit fibrillation at high protein concentrations.

Taru Deva, Nikolai Lorenzen, Brian S Vad, Steen V Petersen, Ida Thørgersen, Jan J Enghild, Torsten Kristensen, Daniel E Otzen

Center for Insoluble Protein Structures (inSPIN), Interdisciplinary Nanoscience Center (iNANO), Department of Molecular Biology and Genetics, University of Aarhus, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark. taru.deva@gmail.com

Biochimica et biophysica acta 2013 Mar

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Ribosomal protein S6 fibrillates readily at slightly elevated temperatures and acidic pH. We find that S6 fibrillation is retarded rather than favored when the protein concentration is increased above a threshold concentration of around 3.5mg/mL. We name this threshold concentration C(FR), the concentration at which fibrillation is retarded. Our data are consistent with a model in which this inhibition is due to the formation of an off-pathway oligomeric species with native-like secondary structure. The oligomeric species dominates at high protein concentrations but exists in dynamic equilibrium with the monomer so that seeding with fibrils can overrule oligomer formation and favors fibrillation under C(FR) conditions. Thus, fibrillation competes with formation of off-pathway oligomers, probably due to a monomeric conversion step that is required to commit the protein to the fibrillation pathway. The S6 oligomer is resistant to pepsin digestion. We also report that S6 forms different types of fibrils dependent on protein concentration. Our observations highlight the multitude of conformational states available to proteins under destabilizing conditions. Copyright © 2013 Elsevier B.V. All rights reserved.

Citation

Taru Deva, Nikolai Lorenzen, Brian S Vad, Steen V Petersen, Ida Thørgersen, Jan J Enghild, Torsten Kristensen, Daniel E Otzen. Off-pathway aggregation can inhibit fibrillation at high protein concentrations. Biochimica et biophysica acta. 2013 Mar;1834(3):677-87