title: GXXXG-Mediated Parallel and Antiparallel Dimerization of Transmembrane Helices and Its Inhibition by Cholesterol: Single-Pair FRET and 2D IR Studies.
abstract: Small-residue-mediated interhelical packings are ubiquitously found in helical membrane proteins, although their interaction dynamics and lipid dependence remain mostly uncharacterized. We used a single-pair FRET technique to examine the effect of a GXXXG motif on the association of de novo designed (AALALAA)3helices in liposomes. Dimerization occurred with sub-second lifetimes, which was abolished by cholesterol. Utilizing the nearly instantaneous time-resolution of 2D IR spectroscopy, parallel and antiparallel helix associations were identified by vibrational couplings across helices at their interface. Taken together, the data illustrate that the GXXXG motif controls helix packing but still allows for a dynamic and lipid-regulated oligomeric state.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yano
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Kondo
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Department of Chemistry, University of Wisconsin, Madison, WI, 53706, USA.
authors.forename: Tianqi O
authors.initials: TO
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, University of Wisconsin, Madison, WI, 53706, USA.
authors.forename: Jia-Jung
authors.initials: JJ
authors.lastname: Ho
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Oishi
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Fujii
authors.affiliation: Department of Chemistry, University of Wisconsin, Madison, WI, 53706, USA.
authors.forename: Martin T
authors.initials: MT
authors.lastname: Zanni
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Katsumi
authors.initials: K
authors.lastname: Matsuzaki
mesh_terms: D000595:Amino Acid Sequence; D002784:Cholesterol / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D008081:Liposomes / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D013055:Spectrophotometry, Infrared / Q000379:methods
chemical_list: D008081:Liposomes; D010455:Peptides; D002784:Cholesterol
keywords: FRET; GXXXG motif; IR spectroscopy; membrane proteins; single-molecule studies