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Instructor: Ashutosh Tripathy; 962-2362 (Ashutosh_Tripathy@med.unc.edu)

Meeting Dates: Last 3rd of spring semester (3/28/2022-4/27/2022)

Time: Lectures MWF 11:15AM-12:05PM

Location:  GMB 3007; Labs TBA in MacInFac

*Registration limited to 16 students. Cross listed as Chem 735.

Macromolecular Interactions Facility (Mac-In-Fac)

 

This is a practical course that coordinates lectures with experience in the Macromolecular Interactions Facility. Lectures introduce methods for monitoring interactions of macromolecules with other molecules. Labs offer students hands-on experience with major techniques available in the Facility.

 

  • Measurement of binding using Isothermal Titration Calorimetry (ITC). Setting up an ITC experiment, and analyzing data to obtain stoichiometry (N), dissociation constant (Kd), and other thermodynamic parameters. 
  • Analytical Ultracentrifugation (AUC): Sedimentation Equilibrium and Sedimentation Velocity. Setting up an AUC experiment to determine molecular weight and oligomeric state of a protein.  
  • Use of Static and Dynamic Light Scattering to determine molecular weight, root-mean-square radius, hydrodynamic radius, and second virial coefficient of a molecule. Setting up static and dynamic light scattering experiments and analyzing data. 
  • Use of Surface Plasmon Resonance (SPR) to determine bimolecular interactions. Setting up a SPR experiment, and analyzing data to obtain on and off rates, and binding constants. 
  • Time permitting – Overview of Fluorescence, Circular Dichroism, Micro-Scale Thermophoresis (MST), nanoDSF, and Octet techniques.