Macromolecular crowding perturbs protein refolding kinetics: implications for folding inside the cell.
We have studied the effects of macromolecular crowding on protein folding kinetics by studying the oxidative refolding of hen lysozyme in the absence and presence of high concentrations of bovine serum albumin and Ficoll 70. The heterogeneity characteristic of the lysozyme refolding process is preserved under crowded conditions. This, together with the observation that the refolding intermediates that accumulate to significant levels are very similar in the absence and presence of Ficoll, suggests that crowding does not alter substantially the energetics of the protein folding reaction. However, the presence of high concentrations of macromolecules results in the acceleration of the fast track of the refolding process whereas the slow track is substantially retarded. The results can be explained by preferential excluded volume stabilization of compact states relative to more unfolded states, and suggest that, relative to dilute solutions, the rates of many protein folding processes are likely to be altered under conditions that more closely resemble the intracellular environment.
Oxford Centre for Molecular Sciences, New Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QT, UK.
B
B
van den Berg
R
R
Wain
C M
CM
Dobson
R J
RJ
Ellis
D005362:Ficoll / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008956:Models, Chemical; D009113:Muramidase / Q000378:metabolism*; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D017510:Protein Folding; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D046911:Macromolecular Substances; D005362:Ficoll; D012710:Serum Albumin, Bovine; D009113:Muramidase