title: Viscoelastic study of the mechanical unfolding of a protein by AFM.
abstract: We have applied a dynamic force modulation technique to the mechanical unfolding of a homopolymer of immunoglobulin (Ig) domains from titin, (C47S C63S I27)5, [(I27)5] to determine the viscoelastic response of single protein molecules as a function of extension. Both the stiffness and the friction of the homopolymer system show a sudden decrease when a protein domain unfolds. The decrease in measured friction suggests that the system is dominated by the internal friction of the (I27)5 molecule and not solvent friction. In the stiffness-extension spectrum we detected an abrupt feature before each unfolding event, the amplitude of which decreased with each consecutive unfolding event. We propose that these features are a clear indication of the formation of the known unfolding intermediate of I27, which has been observed previously in constant velocity unfolding experiments. This simple force modulation AFM technique promises to be a very useful addition to constant velocity experiments providing detailed viscoelastic characterization of single molecules under extension.
authors.forename: Masaru
authors.initials: M
authors.lastname: Kawakami
authors.forename: Katherine
authors.initials: K
authors.lastname: Byrne
authors.forename: David J
authors.initials: DJ
authors.lastname: Brockwell
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
authors.forename: D Alastair
authors.initials: DA
authors.lastname: Smith
mesh_terms: D000818:Animals; D064211:Connectin; D004548:Elasticity; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods; D009124:Muscle Proteins / Q000737:chemistry* / Q000648:ultrastructure; D017510:Protein Folding; D011494:Protein Kinases / Q000737:chemistry* / Q000648:ultrastructure; D017434:Protein Structure, Tertiary / Q000502:physiology; D014783:Viscosity
chemical_list: D064211:Connectin; D007136:Immunoglobulins; D009124:Muscle Proteins; C577285:TTN protein, human; D011494:Protein Kinases