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Understanding the bottlenecks in membrane protein biogenesis

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Title Understanding the bottlenecks in membrane protein biogenesis
Period 12 / 2010 - 05 / 2016
Status Current
Research number OND1344765
Data Supplier NWO

Abstract

The major hurdle in the analysis and exploitation of complex membrane proteins is the overexpression of such proteins in a functional state. Although the mechanisms underlying synthesis, folding, membrane insertion and post-translational modification are largely known, the kinetics, dynamics and location of the individual steps of membrane protein biogenesis are poorly documented. We aim to determine the bottlenecks and roadblocks in membrane protein synthesis in order to obtain a mechanistic understanding of the membrane protein biogenesis pathway in Lactocococcus lactis. We will quantify the intermediates of membrane protein synthesis and determine their location and dynamics in the cell, using engineered and evolved strains that produce the proteins at improved levels and in a functionally competent state. We will use cutting edge single-molecule fluorescence methods (incl. super-resolution microscopy), state-of-the-art molecular biology (incl. deep sequencing of RNA), and biochemistry tools to unravel the intricacies of the membrane protein biogenesis pathway in L. lactis.

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Project leader Prof.dr. J. Kok
Project leader Prof.dr. A.M. van Oijen
Project leader Prof.dr. B. Poolman

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