Molecular Biotechnology

, Volume 58, Issue 4, pp 299–310 | Cite as

Lactococcus lactis is an Efficient Expression System for Mammalian Membrane Proteins Involved in Liver Detoxification, CYP3A4, and MGST1

  • Sana Bakari
  • Mehdi Lembrouk
  • Laura Sourd
  • Fares Ousalem
  • François André
  • Stéphane Orlowski
  • Marcel Delaforge
  • Annie Frelet-Barrand
Original Paper

Abstract

Despite the great importance of human membrane proteins involved in detoxification mechanisms, their wide use for biochemical approaches is still hampered by several technical difficulties considering eukaryotic protein expression in order to obtain the large amounts of protein required for functional and/or structural studies. Lactococcus lactis has emerged recently as an alternative heterologous expression system to Escherichia coli for proteins that are difficult to express. The aim of this work was to check its ability to express mammalian membrane proteins involved in liver detoxification, i.e., CYP3A4 and two isoforms of MGST1 (rat and human). Genes were cloned using two different strategies, i.e., classical or Gateway-compatible cloning, and we checked the possible influence of two affinity tags (6×-His-tag and Strep-tag II). Interestingly, all proteins could be successfully expressed in L. lactis at higher yields than those previously obtained for these proteins with classical expression systems (E. coli, Saccharomyces cerevisiae) or those of other eukaryotic membrane proteins expressed in L. lactis. In addition, rMGST1 was fairly active after expression in L. lactis. This study highlights L. lactis as an attractive system for efficient expression of mammalian detoxification membrane proteins at levels compatible with further functional and structural studies.

Keywords

Lactococcus lactis Membrane proteins Cytochrome P450 Microsomal glutathione S-transferase Gateway 

Supplementary material

12033_2016_9928_MOESM1_ESM.pdf (289 kb)
Supplementary material 1 (PDF 289 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Sana Bakari
    • 1
  • Mehdi Lembrouk
    • 1
  • Laura Sourd
    • 1
  • Fares Ousalem
    • 1
  • François André
    • 1
  • Stéphane Orlowski
    • 1
  • Marcel Delaforge
    • 1
  • Annie Frelet-Barrand
    • 1
    • 2
  1. 1.Institute of Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-SudUniversité Paris-SaclayGif-sur-Yvette CedexFrance
  2. 2.Institute FEMTO-ST, UMR6174 CNRS-Université de Franche-ComtéBesançon CedexFrance

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