Ephemeral states in protein folding under force captured with a magnetic tweezers design

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 116; no. 16; pp. 7873 - 7878
• Main Authors: Tapia-Rojo, Rafael, Eckels, Edward C., Fernández, Julio M.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 16.04.2019
• Subjects: Biological Sciences; Dependence; Design; Disks; Equipment Design; Folding; Force measurement; Magnetic Fields; Magnetic tape; Mechanical Phenomena; Microscopy, Atomic Force; Physical Sciences; Protein Folding; Protein L; Proteins; Proteins - chemistry; Spectroscopy; Spectrum Analysis - instrumentation; Spectrum Analysis - methods; Talin; Variation; Very high frequencies; molten globule state; protein folding; magnetic tape head; protein mechanics; dynamic force spectroscopy
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1821284116

Magnetic tape heads are ubiquitously used to read and record on magnetic tapes in technologies as diverse as old VHS tapes, modern hard-drive disks, or magnetic bands on credit cards. Their design highlights the ability to convert electric signals into fluctuations of the magnetic field at very high frequencies, which is essential for the high-density storage demanded nowadays. Here, we twist this conventional use of tape heads to implement one in a magnetic tweezers design, which offers the unique capability of changing the force with a bandwidth of ∼10 kHz. We calibrate our instrument by developing an analytical expression that predicts the magnetic force acting on a superparamagnetic bead based on the Karlqvist approximation of the magnetic field created by a tape head. This theory is validated by measuring the force dependence of protein L unfolding/folding step sizes and the folding properties of the R3 talin domain. We demonstrate the potential of our instrument by carrying out millisecond-long quenches to capture the formation of the ephemeral molten globule state in protein L, which has never been observed before. Our instrument provides the capability of interrogating individual molecules under fast-changing forces with a control and resolution below a fraction of a piconewton, opening a range of force spectroscopy protocols to study protein dynamics under force.