title: Folding myoglobin within a sol-gel glass: protein folding constrained to a small volume.
abstract: The unfolding and refolding reaction of myoglobin was examined in solution and within a porous silica sol-gel glass. The sol-gel pores constrain the protein to a volume that is the same size and shape as the folded native state accompanied by a few layers of water solvation. Denaturants such as low pH buffers can be diffused through the gel pores to the protein to initiate unfolding and refolding. Acid-induced unfolding was hindered by the steric constraints imposed by the gel pores such that more denaturing conditions were required within the gel than in solution to create the unfolded state. No new folding intermediates were observed. Refolding of myoglobin was not complete in millimolar pH 7 buffer alone. Addition of 25% glycerol to the pH 7 buffer resulted in nearly complete refolding, and the use of 1 M phosphate buffer resulted in complete refolding. The role of this cosolvent and salt in disrupting the ordered water surrounding the protein within the gel is discussed in light of the Hofmeister series and entropic trapping via a diminished hydrophobic effect within the gel. These results are consistent with the premises of folding models in which secondary and tertiary structures are considered to form within a compact conformation of the protein backbone.
authors.affiliation: Department of Chemistry, Concordia College, Moorhead, Minnesota 56562, USA. epeterso@cord.edu
authors.forename: Eric S
authors.initials: ES
authors.lastname: Peterson
authors.forename: Emma F
authors.initials: EF
authors.lastname: Leonard
authors.forename: Jocelyn A
authors.initials: JA
authors.lastname: Foulke
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Oliff
authors.forename: Rosanne D
authors.initials: RD
authors.lastname: Salisbury
authors.forename: David Y
authors.initials: DY
authors.lastname: Kim
mesh_terms: D003198:Computer Simulation; D005898:Glass / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D009211:Myoglobin / Q000737:chemistry*; D044367:Phase Transition; D011487:Protein Conformation; D017510:Protein Folding; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D009211:Myoglobin; D012822:Silicon Dioxide