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

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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.

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Acknowledgments

We are grateful to Denis Pompon (LISBP, Toulouse, France) for his gift of the pYEDP60-CYP3A4 plasmid, Ralf Morgenstern (IEM, Karolinska Institute, Sweden) for cDNA encoding rMGST1 and for fruitful exchanges, to Isabelle de Waziers (Inserm U775, Paris V, France) for CYP3A4-specific antibody, and to François Parcy (LPCV, CEA Grenoble, France) for pBS-RfA plasmid. We thank Ludovic Robillard for technical assistance and Elizabeth Kish-Perrin for linguistic revisions of the manuscript. This work has been supported by the Region Ile de France (DIM SeNt, PhD fellowship to SB AAP2010-3-10T6).

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  1. Annie Frelet-Barrand

    Present address: Institute FEMTO-ST, UMR6174 CNRS-Université de Franche-Comté, 25044, Besançon Cedex, France

Authors and Affiliations

  1. Institute of Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Université Paris-Saclay, 91198, Gif-sur-Yvette Cedex, France

    Sana Bakari, Mehdi Lembrouk, Laura Sourd, Fares Ousalem, François André, Stéphane Orlowski, Marcel Delaforge & Annie Frelet-Barrand

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  1. Sana Bakari
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  2. Mehdi Lembrouk
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Correspondence to Annie Frelet-Barrand.

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Sana Bakari and Mehdi Lembrouk have contributed equally to this work.

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Bakari, S., Lembrouk, M., Sourd, L. et al. Lactococcus lactis is an Efficient Expression System for Mammalian Membrane Proteins Involved in Liver Detoxification, CYP3A4, and MGST1. Mol Biotechnol 58, 299–310 (2016). https://doi.org/10.1007/s12033-016-9928-z

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