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AKB: Biologische Physik

AKB 30: Poster Session I

AKB 30.20: Poster

Montag, 27. März 2006, 15:30–18:00, P1

Single-molecule enzyme kinetics in picoliter confined volumes — •Wolfgang Staroske, Fedor Malik, and Petra Schwille — BIOTEC/TU Dresden, Tatzberg 47-51, 01307 Dresden, Germany

Studying the activity of few or single enzyme molecules under physiological conditions is an exciting task for quantitative biochemistry and biophysics. In particular, the role of confinement in small compartments needs to be determined to better understand these molecules in their cellular environment. We show that using microfluidic devices, homodisperse buffer droplets can be easily prepared in variable sizes, containing very low quantities of enzyme down to the limit of one enzyme molecule per droplet. By observing the fluorescence signal as a measure for substrate or product concentration in a temperature-controlled incubation chamber on an inverted microscope, enzyme activity can be precisely determined. Because of the exact and easy replication of the droplets, it is possible to observe many single enzyme kinetics at once. As a perspective, we aim to elucidate the kinetics of the 20S-Proteasome as a protein degradation machinery.