title: Single-molecule adhesion forces and attachment lifetimes of myosin-I phosphoinositide interactions.
abstract: Phosphoinositides regulate the activities and localization of many cytoskeletal proteins involved in crucial biological processes, including membrane-cytoskeleton adhesion. Yet little is known about the mechanics of protein-phosphoinositide interactions, or about the membrane-attachment mechanics of any peripheral membrane proteins. Myosin-Ic (myo1c) is a molecular motor that links membranes to the cytoskeleton via phosphoinositide binding, so it is particularly important to understand the mechanics of its membrane attachment. We used optical tweezers to measure the strength and attachment lifetime of single myo1c molecules as they bind beads coated with a bilayer of 2% phosphatidylinositol 4,5-bisphosphate and 98% phosphatidylcholine. Adhesion forces measured under ramp-load ranged between 5.5 and 16 pN at loading rates between 250 and 1800 pN/s. Dissociation rates increased linearly with constant force (0.3-2.5 pN), with rates exceeding 360 s(-1) at 2.5 pN. Attachment lifetimes calculated from adhesion force measurements were loading-rate-dependent, suggesting nonadiabatic behavior during pulling. The adhesion forces of myo1c with phosphoinositides are greater than the motors stall forces and are within twofold of the force required to extract a lipid molecule from the membrane. However, attachment durations are short-lived, suggesting that phosphoinositides alone do not provide the mechanical stability required to anchor myo1c to membranes during multiple ATPase cycles.
authors.affiliation: Pennsylvania Muscle Institute and Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
authors.forename: Serapion
authors.initials: S
authors.lastname: Pyrpassopoulos
authors.forename: Henry
authors.initials: H
authors.lastname: Shuman
authors.forename: E Michael
authors.initials: EM
authors.lastname: Ostap
mesh_terms: D000268:Adhesiveness; D000818:Animals; D001696:Biomechanical Phenomena; D019921:Biotinylation; D008051:Lipid Bilayers / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D024461:Myosin Type I; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D011485:Protein Binding; D013997:Time Factors
chemical_list: D008051:Lipid Bilayers; C470422:Myo1c protein, mouse; D019269:Phosphatidylinositol 4,5-Diphosphate; D024461:Myosin Type I; D009218:Myosins