title: High-fidelity protein targeting into membrane lipid microdomains in living cells.
abstract: Lipid analogues carrying three nitrilotriacetic acid (tris-NTA) head groups were developed for the selective targeting of His-tagged proteins into liquid ordered (lo ) or liquid disordered (ld ) lipid phases. Strong partitioning into the lo phase of His-tagged proteins bound to tris-NTA conjugated to saturated alkyl chains (tris-NTA DODA) was achieved, while tris-NTA conjugated to an unsaturated alkyl chain (tris-NTA SOA) predominantly resided in the ld phase. Interestingly, His-tag-mediated lipid crosslinking turned out to be required for efficient targeting into the lo phase by tris-NTA DODA. Robust partitioning into lo phases was confirmed by using viral lipid mixtures and giant plasma membrane vesicles. Moreover, efficient protein targeting into lo and ld domains within the plasma membrane of living cells was demonstrated by single-molecule tracking, thus establishing a highly generic approach for exploring lipid microdomains in situ.
authors.affiliation: Division of Biophysics, Department of Biology, University of Osnabrück, Barbarastrasse 11, 49076 Osnabrück (Germany) http://www.biophysik.uni-osnabrueck.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Beutel
authors.forename: Jörg
authors.initials: J
authors.lastname: Nikolaus
authors.forename: Oliver
authors.initials: O
authors.lastname: Birkholz
authors.forename: Changjiang
authors.initials: C
authors.lastname: You
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmidt
authors.forename: Andreas
authors.initials: A
authors.lastname: Herrmann
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
mesh_terms: D000085:Acetates / Q000737:chemistry*; D004058:Diffusion; D006367:HeLa Cells; D006639:Histidine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism*; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D009574:Nitro Compounds / Q000737:chemistry / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000378:metabolism
chemical_list: D000085:Acetates; C471213:His-His-His-His-His-His; D058619:Maltose-Binding Proteins; D009574:Nitro Compounds; D009842:Oligopeptides; D011506:Proteins; D011993:Recombinant Fusion Proteins; D053835:Unilamellar Liposomes; D006639:Histidine; D009571:Nitrilotriacetic Acid
keywords: lipid rafts; lipid-phase separation; multivalent chelators; proteins; single-molecule studies