title: Slowing down downhill folding: a three-probe study.
abstract: The mutant Tyr22Trp/Glu33Tyr/Gly46Ala/Gly48Ala of lambda repressor fragment lambda(6-85) was previously assigned as an incipient downhill folder. We slow down its folding in a cryogenic water-ethylene-glycol solvent (-18 to -28 degrees C). The refolding kinetics are probed by small-angle x-ray scattering, circular dichroism, and fluorescence to measure the radius of gyration, the average secondary structure content, and the native packing around the single tryptophan residue. The main resolved kinetic phase of the mutant is probe independent and faster than the main phase observed for the pseudo-wild-type. Excess helical structure formed early on by the mutant may reduce the formation of turns and prevent the formation of compact misfolded states, speeding up the overall folding process. Extrapolation of our main cryogenic folding phase and previous T-jump measurements to 37 degrees C yields nearly the same refolding rate as extrapolated by Oas and co-workers from NMR line-shape data. Taken together, all the data consistently indicate a folding speed limit of approximately 4.5 micros for this fast folder.
authors: Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors: Seung Joong
authors: SJ
authors: Kim
authors: Yoshitaka
authors: Y
authors: Matsumura
authors: Charles
authors: C
authors: Dumont
authors: Hiroshi
authors: H
authors: Kihara
authors: Martin
authors: M
authors: Gruebele
mesh_terms: D002942:Circular Dichroism; D005453:Fluorescence; D019791:Guanidine / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012542:Scattering, Radiation; D013696:Temperature; D013997:Time Factors; D054334:Viral Regulatory and Accessory Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014965:X-Rays
chemical_list: D012097:Repressor Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins; D019791:Guanidine