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'Pre steady-state kinetics of redox metalloenzymes using ultrafast mixing techniques' with Dr Peter-Leon Hagedoorn from the University of Delft

Image: Pre steady-state kinetics of redox metalloenzymes
Pre steady-state kinetics of redox metalloenzymes

Date: 16 February 2018   Time: 12:30 - 13:30

'Pre steady-state kinetics of redox metalloenzymes using ultrafast mixing techniques'
Peter Leon Hagedoorn, Batoul Srour, Marc J.F. Strampraad, and Wilfred R. Hagen
Delft University of Technology, Department of Biotechnology, Van der Maasweg 9, 2629HZ, Delft, The Netherlands

Understanding the molecular mechanism of enzyme catalysis requires detailed structural and mechanistic information that can only be obtained through a combination of techniques and experiments. Pre-steady state, or rapid kinetics techniques, such as stopped-flow UV-vis absorbance of fluorescence spectroscopy are highly important tools to determine rate constants of individual steps in a catalytic mechanism, and to identify intermediates that will help us to understand the mechanism. The dead time of a conventional stopped flow instrument is approximately 1 ms, which does not allow the observation of intermediates with a shorter lifetime. By using a specially designed four-jet tangential micromixer we have been able to construct a continuous flow UV-vis spectroscopic device, called the Nanospec, with a dead time of 4 ?s.1 And, using the same mixer, a freeze quench instrument called the Microsecond freeze Hyper Quenching device (MHQ) with a dead time of 80 ?s.2 The working principle of these instruments and data on cytochrome c refolding kinetics will be presented as well as preliminary data on particular enzymes.

Location:  Laws 2.10, Mile End Campus
Arranged by::  Peter-Leon Hagedoorn Group
Contact:  Dr Maxie Roessler
Email:  p.l.hagedoorn@tudelft.nl