Sellers - Takagi - 2019

Laboratory of Molecular Physiology (LMP)
Mentor Name
James R. Sellers, Ph.D.
Yasuharu “Harry” Takagi, Ph.D.
Mentor Telephone
(301) 496-6887
Laboratory And Project Description

The Laboratory of Molecular Physiology (LMP) focuses on the understanding of cytoskeletal proteins, specifically myosins, a molecular motor that converts chemical energy (ATP) to mechanical output (work/force/displacement). To understand the detailed mechanism of the myosin superfamily, we undertake a multidisciplinary approach, including techniques from molecular biology, cell biology, biophysics, biochemistry and engineering.
Recently in our lab, we have used a couple of single molecule techniques: (1) single molecule total internal reflection fluorescence (TIRF) microscopy and (2) single molecule optical trapping microscopy. Both techniques were used to determine information that could only be interrogated using single molecule approaches such as step-size, power-stroke size and stall force, but also information that could be compared to bulk solution studies such as the kinetics, or transition rates between specific states, of these motor proteins.
The projects in the LMP available to the Summer 2017 BESIP interns includes the following:

  • Molecular biology and expression of different mutants of the myosins we study in the lab using the Sf9/baculovirus expression system;
  • Use of the stopped flow transient kinetic apparatus to characterize kinetic parameters;
  • Use of the optical trap or TIRF microscope to characterize the single molecule mechanics/kinetics of various myosins;
  • Fixed/live cell imaging of myosins in a variety of cell lines using fluorescence microscopy techniques; and
  • Instrumentation design using state of the art lasers, optical components and detectors.