title: Curvature enhances binding and aggregation of huntingtin at lipid membranes.
abstract: Huntington disease (HD) is a genetic neurodegenerative disease caused by an expanded polyglutamine (polyQ) domain in the first exon of the huntingtin (Htt) protein, facilitating its aggregation. Htt interacts with a variety of membraneous structures within the cell, and the first 17 amino acids (Nt17) of Htt directly flanking the polyQ domain comprise an amphiphathic α-helix (AH) lipid-binding domain. AHs are also known to detect membrane curvature. To determine if Htt exon 1 preferentially binds curved membranes, in situ atomic force microscopy (AFM) studies were performed. Supported lipid bilayers are commonly used as model membranes for AFM studies of protein aggregation. However, these supported bilayers usually lack curvature. By forming a bilayer on top of silica nanobeads (50 ± 10 nm) deposited on a silicon substrate, model supported lipid bilayers with flat and curved regions were developed for AFM studies. The presence of the bilayer over the beads was validated by continual imaging of the formation of the bilayer, height measurements, and spatially resolved mechanical measurements of the resulting bilayer using scanning probe acceleration microscopy. Interpretation of this data was facilitated by numerical simulations of the entire imaging process. The curved supported bilayers associated with the beads were found to be more compliant than flat supported bilayers, consistent with the altered packing density of lipids caused by the induced curvature. This model bilayer system was exposed to a synthetic truncated Htt exon 1 peptide (Nt17Q35P10KK), and this peptide preferentially accumulated on curved membranes, consistent with the ability of AHs to sense membrane curvature.
authors.affiliation: The C. Eugene Bennett Department of Chemistry, West Virginia University , Morgantown, West Virginia 26505, United States.
authors.forename: Maxmore
authors.initials: M
authors.lastname: Chaibva
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Burke
authors.forename: Justin
authors.initials: J
authors.lastname: Legleiter
mesh_terms: D000071058:Huntingtin Protein; D008051:Lipid Bilayers / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D053758:Nanoparticles; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D008051:Lipid Bilayers; D009419:Nerve Tissue Proteins