Abstract
Heterologous expression is a landmark technique for studying a protein itself or its effect on the expression host, in which membrane-embedded proteins are a common choice. Yet, the impact of inserting a foreign protein to the lipid environment of host membranes, has never been addressed. Here we demonstrated that heterologous expression of the Artemia franciscana adenine nucleotide translocase (ANT) in yeasts altered lipidomic composition of their inner mitochondrial membranes. Along with this, activities of complex II, IV and ATP synthase, all membrane-embedded components, were significantly decreased while their expression levels remained unaffected. Although the results represent an individual case of expressing a crustacean protein in yeast inner mitochondrial membranes, it cannot be excluded that host lipidome alterations is a more widespread epiphenomenon, potentially biasing heterologous expression experiments. Finally, our results raise the possibility that not only lipids modulate protein function, but also membrane-embedded proteins modulate lipid composition, thus revealing a reciprocal mode of regulation for these two biomolecular entities.
Highlights
Heterologous expression is a landmark technique for studying a protein itself or its effect on the expression host, in which membrane-embedded proteins are a common choice
That number increases to 90 statistically significant lipid molecular species when compared to isolated mitochondria (Fig. 1B), and increases further to 101 lipid molecular species in isolated mitoplasts (Fig. 1C)
In the present study we investigated the effect of heterologous expression of ArANT in yeast by examining their total, mitochondrial and mitoplastic lipidomes and interrogating the function of other proteins embedded in the same membrane as the heterologously expressed one
Summary
Heterologous expression is a landmark technique for studying a protein itself or its effect on the expression host, in which membrane-embedded proteins are a common choice. The results represent an individual case of expressing a crustacean protein in yeast inner mitochondrial membranes, it cannot be excluded that host lipidome alterations is a more widespread epiphenomenon, potentially biasing heterologous expression experiments. Ca2+ or pH) and/or is targeted towards a particular compartment, or it is intentionally overexpressed for the purpose of protein purification or it is plasma membrane-targeted for raising ligands or antibodies against it. The latter application is inherently relevant to the pharmaceutical industry, because membrane proteins are the targets of >50% of drugs despite representing only ~1% of total cellular proteins[2]. It is not surprising that a Pubmed search for the string [“heterologous expression” OR “exogenous expression” OR “ectopic expression”]
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