title: Direct observation of downhill folding of lambda-repressor in a microfluidic mixer.
abstract: The protein lambda(6-85) has been implicated in barrierless folding by observations of kinetic relaxation after nanosecond T-jump. In this work we observed folding of this protein after dilution of a high denaturant in an ultrarapid microfluidic mixer at temperatures far below the thermal midpoint. The observations of total intensity and spectral shift of tryptophan fluorescence yielded distinctly different kinetics and activation energies. These results may be explained as diffusion on a low-barrier, one-dimensional, free-energy surface, with different probes having different sensitivities along the reaction coordinate. Additionally, we observed an extremely fast phase within the mixing time that was not observed by T-jump, suggesting that the ensemble of unfolded states populated at high denaturant is distinct from those accessible at high temperature.
authors.affiliation: Department of Physics and Astronomy, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: DeCamp
authors.forename: Athi N
authors.initials: AN
authors.lastname: Naganathan
authors.forename: Steven A
authors.initials: SA
authors.lastname: Waldauer
authors.forename: Olgica
authors.initials: O
authors.lastname: Bakajin
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Lapidus
mesh_terms: D007700:Kinetics; D046210:Microfluidic Analytical Techniques; D009154:Mutation; D011489:Protein Denaturation; D017510:Protein Folding; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics; D054334:Viral Regulatory and Accessory Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D012097:Repressor Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins