Functional expression of mammalian receptors and membrane channels in different cells.
In native tissues, the majority of medically important membrane proteins is only present at low concentrations, making their overexpression in recombinant systems a prerequisite for structural studies. Here, we explore the commonly used eukaryotic expression systems-yeast, baculovirus/insect cells (Sf9) and Semliki Forest Virus (SFV)/mammalian cells-for the expression of seven different eukaryotic membrane proteins from a variety of protein families. The expression levels, quality, biological activity, localization and solubility of all expressed proteins are compared in order to identify the advantages of one system over the other. SFV-transfected mammalian cell lines provide the closest to native environment for the expression of mammalian membrane proteins, and they exhibited the best overall performance. But depending on the protein, baculovirus-infected Sf9 cells performed almost as well as mammalian cells. The lowest expression levels for the proteins tested here were obtained in yeast.
Eifler
Nora
N
Maurice E. Müller Institute for Microscopy, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
Duckely
Myriam
M
Sumanovski
Lazar T
LT
Egan
Terrance M
TM
Oksche
Alexander
A
Konopka
James B
JB
Lüthi
Anita
A
Engel
Andreas
A
Werten
Paul J L
PJ
D000818:Animals; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D008565:Membrane Proteins; D011994:Recombinant Proteins