title: Segmental Deuteration of α-Synuclein for Neutron Reflectometry on Tethered Bilayers.
abstract: Neutron reflectometry (NR) is uniquely suited for studying protein interaction with phospholipid bilayers along the bilayer normal on an angstrom scale. However, NR on its own cannot discern specific membrane-bound regions due to a lack of scattering contrast within a protein. Here we report the successful coupling of native chemical ligation (NCL) and NR to study α-synuclein (α-syn), a membrane-binding neuronal protein central in Parkinson's disease. Two α-syn variants were generated where either the first 86 or last 54 residues are deuterated, allowing for region-specific contrast within the protein and the identification of membrane interacting residues by NR. Residues 1-86 are positioned at the hydrocarbon/headgroup interface of the outer leaflet, whereas the density distribution of the 54 C-terminal residues ranges from the hydrocarbon region to the aqueous environment. Coupling of NCL and NR should have broad utility in studies of membrane protein folding.
authors.affiliation: Laboratory of Protein Conformation and Dynamics, National Heart, Lung, and Blood Institute, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: Department of Physics, Carnegie Mellon University , Pittsburgh, Pennsylvania 15213, United States.|National Institute of Standards and Technology , Gaithersburg, Maryland 20899, United States.
authors.forename: Frank
authors.initials: F
authors.lastname: Heinrich
authors.affiliation: Laboratory of Protein Conformation and Dynamics, National Heart, Lung, and Blood Institute, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors.forename: Ryan P
authors.initials: RP
authors.lastname: McGlinchey
authors.affiliation: Laboratory of Protein Conformation and Dynamics, National Heart, Lung, and Blood Institute, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors.forename: James M
authors.initials: JM
authors.lastname: Gruschus
authors.affiliation: Laboratory of Protein Conformation and Dynamics, National Heart, Lung, and Blood Institute, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors.forename: Jennifer C
authors.identifier: http://orcid.org/0000-0003-0506-8349
authors.initials: JC
authors.lastname: Lee