Femtomole mixer for microsecond kinetic studies of protein folding

被引:119
作者
Hertzog, DE
Michalet, X
Jäger, M
Kong, XX
Santiago, JG
Weiss, S
Bakajin, O [1 ]
机构
[1] Stanford Univ, Dept Mech Engn, Stanford, CA 94305 USA
[2] Lawrence Livermore Natl Lab, Dept Chem & Mat Sci, Livermore, CA 94550 USA
[3] Univ Calif Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90095 USA
关键词
D O I
10.1021/ac048661s
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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.
引用
收藏
页码:7169 / 7178
页数:10
相关论文
共 51 条
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White F. M., 1974, VISCOUS FLUID FLOW