title: Combining single-molecule optical trapping and small-angle x-ray scattering measurements to compute the persistence length of a protein ER/K alpha-helix.
abstract: A relatively unknown protein structure motif forms stable isolated single alpha-helices, termed ER/K alpha-helices, in a wide variety of proteins and has been shown to be essential for the function of some molecular motors. The flexibility of the ER/K alpha-helix determines whether it behaves as a force transducer, rigid spacer, or flexible linker in proteins. In this study, we quantify this flexibility in terms of persistence length, namely the length scale over which it is rigid. We use single-molecule optical trapping and small-angle x-ray scattering, combined with Monte Carlo simulations to demonstrate that the Kelch ER/K alpha-helix behaves as a wormlike chain with a persistence length of approximately 15 nm or approximately 28 turns of alpha-helix. The ER/K alpha-helix length in proteins varies from 3 to 60 nm, with a median length of approximately 5 nm. Knowledge of its persistence length enables us to define its function as a rigid spacer in a translation initiation factor, as a force transducer in the mechanoenzyme myosin VI, and as a flexible spacer in the Kelch-motif-containing protein.
authors: Department of Biochemistry, Stanford University, Stanford, California, USA.
authors: S
authors: S
authors: Sivaramakrishnan
authors: J
authors: J
authors: Sung
authors: M
authors: M
authors: Ali
authors: S
authors: S
authors: Doniach
authors: H
authors: H
authors: Flyvbjerg
authors: J A
authors: JA
authors: Spudich
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D009010:Monte Carlo Method; D052898:Optical Tweezers; D015202:Protein Engineering; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053838:Scattering, Small Angle; D013552:Swine; D013696:Temperature; D014961:X-Ray Diffraction
chemical_list: D011506:Proteins