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Wiley, Protein Science, 12(14), p. 3048-3056, 2005

DOI: 10.1110/ps.051689905

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Functional expression of eukaryotic membrane proteins inLactococcus lactis

Journal article published in 2005 by Magnus Monné, Ka Wai Chan, Dirk-Jan Slotboom, Edmund R. S. Kunji ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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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.