Perturbation of the yeast mitochondrial lipidome and associated membrane proteins following heterologous expression of Artemia-ANT

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.

Mitochondrial Phospholipid Homeostasis Is Regulated by the i-AAA Protease PaIAP and Affects Organismic Aging

“…The abundance of DAG was found to be rather high with 25% of the analyzed PLs, possibly indicating a co-purification of mitochondria-attached ER or lipid droplets. Nevertheless, the amount of DAG is comparable to previous observations in highly purified yeast mitochondria [46], which is why we expected the quality of the isolated mitochondrial fractions to be sufficient for further experiments. Surprisingly, we detected only low amounts of the mitochondrial signature PL cardiolipin (1.5%), which, in other species like yeast, mammals or plants, represents 10-15% of mitochondrial PLs [47].…”

An essential role for cardiolipin in the stability and function of the mitochondrial calcium uniporter

“…1 B ) ( 17 ) and are thus expected to influence membrane protein function. Previous work has shown that heterologous expression of membrane proteins in yeast can itself perturb the phospholipid composition ( 20 ), which can confound our study. Therefore, we first tested the effect of expressing MCU on the phospholipid composition of WT yeast cells.…”

Faculty Opinions recommendation of An essential role for cardiolipin in the stability and function of the mitochondrial calcium uniporter.

“…1B) (17) and are thus expected to influence membrane protein function. Previous work has shown that heterologous expression of membrane proteins in yeast can itself perturb the phospholipid composition (20), which can confound our study. Therefore, we first tested the effect of expressing MCU on the phospholipid composition of WT yeast cells.…”