High Hydrostatic Pressure Can Reverse Aggregation of Protein Folding Intermediates and Facilitate Acquisition of Native Structure

• Published in: Biochemistry (Easton) Vol. 37; no. 17; pp. 6132 - 6135
• Main Authors: Gorovits, Boris M, Horowitz, Paul M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 28-04-1998
• Subjects: Buffers; Hydrostatic Pressure; Protein Denaturation; Protein Folding; Protein Structure, Secondary; Protein Structure, Tertiary; Spectrometry, Fluorescence; Thiosulfate Sulfurtransferase - chemistry; Urea
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi9730137

The present work demonstrates that high hydrostatic pressure can increase protein folding by reducing nonspecific aggregation. Protein aggregation is one of the main side reactions that competes with protein folding, and it typically results from interactions among partially folded intermediates. It is known that oligomeric proteins can be dissociated by the application of high hydrostatic pressure. Since protein aggregates can be described as nonspecific protein oligomers, it can be predicted that they can be completely or partially dissociated by pressure. The enzyme rhodanese is prone to slow aggregation in 3.9 M urea, and it is widely used as a model for the folding of a protein which readily aggregates. In the present study, it was demonstrated that this aggregation process could be completely reversed under high hydrostatic pressure. Release of the pressure led to renewed protein aggregation. In addition, it was demonstrated that refolding of urea-denatured rhodanese at 2 kbar pressure led to an increased yield of the native enzyme. The final recovery was increased up to ∼25% in contrast to ∼5% recovery observed under ambient pressure. The recovery can be further increased in the presence of 4 M glycerol, where 56% of the protein was recovered by treatment with high pressure. These observations suggest that some protein aggregation can be limited without the use of chemical additives, and they show that the pressures needed to maintain solubility are considerably less than those typically required for dissociation of specific oligomers and unfolding of polypeptide chains.