The folding pathway of spectrin R17 from experiment and simulation: using experimentally validated MD simulations to characterize States hinted at by experiment.
We present an experimental and computational analysis of the folding pathway of the 17th domain of chicken brain alpha-spectrin, R17. Wild-type R17 folds in a two-state manner and the chevron plot (plot of the logarithm of the observed rate constant against concentration of urea) shows essentially linear folding and unfolding arms. A number of mutant proteins, however, show a change in slope of the unfolding arm at high concentration of denaturant, hinting at complexity in the folding landscape. Through a combination of mutational studies and high temperature molecular dynamics simulations we show that the folding of R17 can be described by a model with two sequential transition states separated by an intermediate species. The rate limiting transition state for folding in water has been characterized both through experimental Phi-value analysis and by simulation. In contrast, a detailed analysis of the transition state predicted to dominate under highly denaturing conditions is only possible by simulation.
Scott
Kathryn A
KA
MRC Centre for Protein Engineering, University of Cambridge Chemical Laboratory, UK.
Randles
Lucy G
LG
Moran
Stephen J
SJ
Daggett
Valerie
V
Clarke
Jane
J
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002645:Chickens; D003198:Computer Simulation; D008958:Models, Molecular; D009154:Mutation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013049:Spectrin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D013049:Spectrin