Leishmania tarentolae as a Promising Tool for Expressing Polytopic and Multi-Transmembrane Spans Eukaryotic Membrane Proteins: The Case of the ABC Pump ABCG6

Methods Mol Biol. 2016:1432:119-31. doi: 10.1007/978-1-4939-3637-3_8.

Abstract

This chapter includes a practical method of membrane protein production in Leishmania tarentolae cells. We routinely use it to express membrane proteins of the ABC (adenosine triphosphate-binding cassette) family, here exemplified with ABCG6 from L. braziliensis, implicated in phospholipid trafficking and drug efflux. The pLEXSY system used here allows membrane protein production with a mammalian-like N-glycosylation pattern, at high levels and at low costs. Also the effects of an N-terminal truncation of the protein are described. The method is described to allow any kind of membrane protein production.

Keywords: ABC transporters; Drug efflux; Leishmania tarentolae; Membrane protein expression.

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / metabolism*
  • Animals
  • Gene Expression
  • Glycosylation
  • Humans
  • Leishmania / genetics
  • Leishmania / growth & development*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Protein Engineering / methods*
  • Protozoan Proteins / genetics

Substances

  • ATP-Binding Cassette Transporters
  • Membrane Proteins
  • Protozoan Proteins