Folding Dynamics and Pathways of the Trp-Cage Miniproteins

• Published in: Biochemistry (Easton) Vol. 53; no. 38; pp. 6011 - 6021
• Main Authors: Byrne, Aimee, Williams, D. Victoria, Barua, Bipasha, Hagen, Stephen J, Kier, Brandon L, Andersen, Niels H
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 30-09-2014
• Subjects: Magnetic Resonance Spectroscopy; Mutation; Protein Conformation; Protein Folding
• DOI: 10.1021/bi501021r
• EISSN: 1520-4995
• ISSN: 0006-2960

Using alternate measures of fold stability for a wide variety of Trp-cage mutants has raised the possibility that prior dynamics T-jump measures may not be reporting on complete cage formation for some species. NMR relaxation studies using probes that only achieve large chemical shift difference from unfolded values on complete cage formation indicate slower folding in some but not all cases. Fourteen species have been examined, with cage formation time constants (1/k F) ranging from 0.9–7.5 μs at 300 K. The present study does not change the status of the Trp-cage as a fast folding, essentially two-state system, although it does alter the stage at which this description applies. A diversity of prestructuring events, depending on the specific analogue examined, may appear in the folding scenario, but in all cases, formation of the N-terminal helix is complete either at or before the cage-formation transition state. In contrast, the fold-stabilizing H-bonding interactions of the buried Ser14 side chain and the Arg/Asp salt bridge are post-transition state features on the folding pathway. The study has also found instances in which a [P12W] mutation is fold destabilizing but still serves to accelerate the folding process.