Interaction of the Spo20 membrane-sensor motif with phosphatidic acid and other anionic lipids, and influence of the membrane environment

The yeast protein Spo20 contains a regulatory amphipathic motif that has been suggested to recognize phosphatidic acid, a lipid involved in signal transduction, lipid metabolism and membrane fusion. We have investigated the interaction of the Spo20 amphipathic motif with lipid membranes using a biop...

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Bibliographic Details
Published in:PloS one 2014-11, Vol.9 (11), p.e113484-e113484
Main Authors: Horchani, Habib, de Saint-Jean, Maud, Barelli, Hélène, Antonny, Bruno
Format: Article
Language:English
Subjects:
Acids
Amino Acid Motifs
Binding
Biology and Life Sciences
Cell division
Cell Line
Epithelial Cells - cytology
Epithelial Cells - metabolism
Gene Expression Regulation
Genes, Reporter
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Helices
Humans
Hydrophobic and Hydrophilic Interactions
Inositol
Ligands
Lipid membranes
Lipid metabolism
Lipids
Liposomes
Liposomes - chemistry
Liposomes - metabolism
Localization
Membrane fusion
Membrane proteins
Membranes
Metabolism
Molecular Probes
Molecular Sequence Data
Mutation
Phosphatidic acid
Phosphatidic Acids - metabolism
Phosphatidylethanolamine
Phosphatidylethanolamines - chemistry
Phosphatidylethanolamines - metabolism
Phosphatidylinositol Phosphates - chemistry
Phosphatidylinositol Phosphates - metabolism
Phosphatidylserine
Phosphatidylserines - chemistry
Phosphatidylserines - metabolism
Protein Binding
Protein structure
Proteins
Qb-SNARE Proteins - chemistry
Qb-SNARE Proteins - genetics
Qb-SNARE Proteins - metabolism
Qc-SNARE Proteins - chemistry
Qc-SNARE Proteins - genetics
Qc-SNARE Proteins - metabolism
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins - chemistry
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
Sensors
Signal Transduction
Surface active agents
Yeast
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Summary:The yeast protein Spo20 contains a regulatory amphipathic motif that has been suggested to recognize phosphatidic acid, a lipid involved in signal transduction, lipid metabolism and membrane fusion. We have investigated the interaction of the Spo20 amphipathic motif with lipid membranes using a bioprobe strategy that consists in appending this motif to the end of a long coiled-coil, which can be coupled to a GFP reporter for visualization in cells. The resulting construct is amenable to in vitro and in vivo experiments and allows unbiased comparison between amphipathic helices of different chemistry. In vitro, the Spo20 bioprobe responded to small variations in the amount of phosphatidic acid. However, this response was not specific. The membrane binding of the probe depended on the presence of phosphatidylethanolamine and also integrated the contribution of other anionic lipids, including phosphatidylserine and phosphatidyl-inositol-(4,5)bisphosphate. Inverting the sequence of the Spo20 motif neither affected the ability of the probe to interact with anionic liposomes nor did it modify its cellular localization, making a stereo-specific mode of phosphatidic acid recognition unlikely. Nevertheless, the lipid binding properties and the cellular localization of the Spo20 alpha-helix differed markedly from that of another amphipathic motif, Amphipathic Lipid Packing Sensor (ALPS), suggesting that even in the absence of stereo specific interactions, amphipathic helices can act as subcellular membrane targeting determinants in a cellular context.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0113484