title: An investigation of the effect of membrane curvature on transmembrane-domain dependent protein sorting in lipid bilayers.
abstract: Sorting of membrane proteins within the secretory pathway of eukaryotic cells is a complex process involving discrete sorting signals as well as physico-chemical properties of the transmembrane domain (TMD). Previous work demonstrated that tail-anchored (TA) protein sorting at the interface between the Endoplasmic Reticulum (ER) and the Golgi complex is exquisitely dependent on the length and hydrophobicity of the transmembrane domain, and suggested that an imbalance between TMD length and bilayer thickness (hydrophobic mismatch) could drive long TMD-containing proteins into curved membrane domains, including ER exit sites, with consequent export of the mismatched protein out of the ER. Here, we tested a possible role of curvature in TMD-dependent sorting in a model system consisting of Giant Unilamellar Vesicles (GUVs) from which narrow membrane tubes were pulled by micromanipulation. Fluorescent TA proteins differing in TMD length were incorporated into GUVs of uniform lipid composition or made of total ER lipids, and TMD-dependent sorting and diffusion, as well as the bending rigidity of bilayers made of microsomal lipids, were investigated. Long and short TMD-containing constructs were inserted with similar orientation, diffused equally rapidly in GUVs and in tubes pulled from GUVs, and no difference in their final distribution between planar and curved regions was detected. These results indicate that curvature alone is not sufficient to drive TMD-dependent sorting at the ER-Golgi interface, and set the basis for the investigation of the additional factors that must be required.
authors.affiliation: CNR Institute of Neuroscience and Department of Biotechnology and Translational Medicine; University of Milano; Milano, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Fossati
authors.affiliation: CNRS-Institut Curie; UMR144; Paris, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Goud
authors.affiliation: CNR Institute of Neuroscience and Department of Biotechnology and Translational Medicine; University of Milano; Milano, Italy ; Department of Health Science; University of Catanzaro "Magna Graecia"; Catanzaro, Italy.
authors.forename: Nica
authors.initials: N
authors.lastname: Borgese
authors.affiliation: CNRS-Institut Curie; UMR144; Paris, France.
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Manneville
keywords: bending rigidity; endoplasmic reticulum; giant unilamellar vesicles; hydrophobic mismatch; nanotubes; optical tweezers; tail-anchored proteins