RT Journal Article
SR Electronic
T1 A Magnetic Head Design for Manipulating Single Proteins with Force
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 310060
DO 10.1101/310060
A1 Rafael Tapia-Rojo
A1 Edward C. Eckels
A1 Julio M. Fernandez
YR 2018
UL http://biorxiv.org/content/early/2018/04/28/310060.abstract
AB Magnetic heads are ubiquitously used to record and read on magnetic tapes in technologies as diverse as old cassettes or VHS tapes, modern hard drive disks, or magnetic bands in credit/debit or subway cards. They are designed to convert electric signals into fluctuations on the magnetic field at very high frequencies, crucial for the high density storage which is demanded nowadays. Here, we twist this traditional use of magnetic heads and implement one in a new force spectrometer design, where the magnetic field is used to pull on proteins tethered to superparamagnetic beads. Our instrument offers the same features as magnetic tweezers—intrinsic force-clamp conditions, with accurate control of the force, and intrinsic stability—, but with the novel ability of changing the force instantaneously, which allows to investigate protein dynamics at very short timescales, or under arbitrary force signals. We calibrate our instrument by relying on Karlqvist approximation of the field created by a magnetic head—the first building block of magnetic recording theory—through the force scaling of the unfolding/folding step-sizes of protein L, used as a molecular template. This results in a force range between 0 and 50 pN, when working at distances above 250 μm, and electric currents up to 1 A. We illustrate the potential of our instrument by studying the folding mechanism of protein L upon ultra-fast force quenches. This allows us to describe that, in a short timescale of 50 ms, the unfolded protein L evolves to an ensemble of weak collapsed states, eventually acquiring the native conformation in a timescale of seconds. Our instrumental development provides a unique capability of interrogating individual molecules under fast-changing force signals, which opens a range of novel force spectroscopy schemes of unexplored biological significance.