title: Femtomole mixer for microsecond kinetic studies of protein folding.
abstract: We have developed a microfluidic mixer for studying protein folding and other reactions with a mixing time of 8 mus and sample consumption of femtomoles. This device enables us to access conformational changes under conditions far from equilibrium and at previously inaccessible time scales. In this paper, we discuss the design and optimization of the mixer using modeling of convective diffusion phenomena and a characterization of the mixer performance using microparticle image velocimetry, dye quenching, and Forster resonance energy-transfer (FRET) measurements of single-stranded DNA. We also demonstrate the feasibility of measuring fast protein folding kinetics using FRET with acyl-CoA binding protein.
authors.affiliation: Mechanical Engineering Department, Stanford University, Stanford, California 94305, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Hertzog
authors.forename: Xavier
authors.initials: X
authors.lastname: Michalet
authors.forename: Marcus
authors.initials: M
authors.lastname: Jäger
authors.forename: Xiangxu
authors.initials: X
authors.lastname: Kong
authors.forename: Juan G
authors.initials: JG
authors.lastname: Santiago
authors.forename: Shimon
authors.initials: S
authors.lastname: Weiss
authors.forename: Olgica
authors.initials: O
authors.lastname: Bakajin
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000032:analysis; D004268:DNA-Binding Proteins / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D007700:Kinetics; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008958:Models, Molecular; D011487:Protein Conformation; D017510:Protein Folding
chemical_list: D000214:Acyl Coenzyme A; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins