Visualizing a one-way protein encounter complex by ultrafast single-molecule mixing.

Gambin, Yann; VanDelinder, Virginia; Ferreon, Allan Chris M; Lemke, Edward A; Groisman, Alex; Deniz, Ashok A · Nat Methods · 2011

basic_science · Level V

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Abstract

We combined rapid microfluidic mixing with single-molecule fluorescence resonance energy transfer to study the folding kinetics of the intrinsically disordered human protein α-synuclein. The time-resolution of 0.2 ms revealed initial collapse of the unfolded protein induced by binding with lipid mimics and subsequent rapid formation of transient structures in the encounter complex. The method also enabled analysis of rapid dissociation and unfolding of weakly bound complexes triggered by massive dilution.

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