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Reference: Monne M, et al. (2005) Functional expression of eukaryotic membrane proteins in Lactococcus lactis. Protein Sci 14(12):3048-56

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Abstract

The overproduction of eukaryotic membrane proteins is a major impediment in their structural and functional characterization. Here we have used the nisin-inducible expression system of Lactococcus lactis for the overproduction of 11 mitochondrial transport proteins from yeast. They were expressed at high levels in a functional state in the cytoplasmic membrane. The results also show that the level of expression is influenced by the N-terminal regions of the transporters. Expression levels were improved >10-fold either by replacing or truncating these regions or by adding lactococcal signal peptides. The observed expression levels are now compatible with a realistic exploration of crystallization conditions. The lactococcal expression system may be used for the high-throughput functional characterization of eukaryotic membrane proteins and structural genomics.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Monne M, Chan KW, Slotboom DJ, Kunji ER
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