title: Protein analysis by time-resolved measurements with an electro-switchable DNA chip.
abstract: Measurements in stationary or mobile phases are fundamental principles in protein analysis. Although the immobilization of molecules on solid supports allows for the parallel analysis of interactions, properties like size or shape are usually inferred from the molecular mobility under the influence of external forces. However, as these principles are mutually exclusive, a comprehensive characterization of proteins usually involves a multi-step workflow. Here we show how these measurement modalities can be reconciled by tethering proteins to a surface via dynamically actuated nanolevers. Short DNA strands, which are switched by alternating electric fields, are employed as capture probes to bind target proteins. By swaying the proteins over nanometre amplitudes and comparing their motional dynamics to a theoretical model, the protein diameter can be quantified with Angström accuracy. Alterations in the tertiary protein structure (folding) and conformational changes are readily detected, and even post-translational modifications are revealed by time-resolved molecular dynamics measurements.
authors.affiliation: Chemistry Department and Walter Schottky Institute, Technische Universität München, Lichtenbergstrasse 4, 85748 Garching, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Langer
authors.forename: Paul A
authors.initials: PA
authors.lastname: Hampel
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Kaiser
authors.forename: Jelena
authors.initials: J
authors.lastname: Knezevic
authors.forename: Thomas
authors.initials: T
authors.lastname: Welte
authors.forename: Valentina
authors.initials: V
authors.lastname: Villa
authors.forename: Makiko
authors.initials: M
authors.lastname: Maruyama
authors.forename: Matej
authors.initials: M
authors.lastname: Svejda
authors.forename: Simone
authors.initials: S
authors.lastname: Jähner
authors.forename: Frank
authors.initials: F
authors.lastname: Fischer
authors.forename: Ralf
authors.initials: R
authors.lastname: Strasser
authors.forename: Ulrich
authors.initials: U
authors.lastname: Rant
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D004560:Electricity; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D006063:Chorionic Gonadotropin; D005656:Fungal Proteins; D007074:Immunoglobulin G; D004247:DNA