AUG 13, 2015 08:00 AM PDT

Analytical ultracentrifugation as a complementary technique for structural analysis of proteins and macromolecular complexes

Speakers
  • Associate Professor, Divisions of Immunobiology & Infectious Diseases, Cincinnati Children's Hospital Medical Center
    Biography
      Andrew Herr, PhD, is an Associate Professor in the Divisions of Immunobiology and Infectious Diseases and the Center for Systems Immunology at Cincinnati Children's Hospital Medical Center. Before his recent move to Cincinnati Children's, he was the Ohio Eminent Scholar in Structural Biology at the University of Cincinnati College of Medicine. Dr. Herr trained in Molecular Biophysics and Structural Biology at Washington University in St. Louis and Caltech before joining the faculty at the University of Cincinnati. Dr. Herr's laboratory uses a combination of analytical ultracentrifugation, X-ray crystallography and a wide array of biophysical techniques to analyze the structure and function of proteins involved in bacterial pathogenesis and the host immune response. For example, they have been studying protein assembly events underlying intercellular adhesion in bacterial biofilms. In this project, AUC data provided the critical clue leading to a novel antimicrobial technology under development for therapeutic use. The Herr lab has used AUC to study proteins that range in size from small polypeptides to massive antibody immune complexes, integral membrane proteins, intrinsically disordered proteins, and complicated assembling systems. They have also worked with biotech and pharma companies to analyze protein samples and help guide formulation decisions. Dr. Herr has authored over 50 published manuscripts and book chapters, with over half of these involving AUC analyses.

    Abstract:
    Analytical ultracentrifugation (AUC) is a powerful technique for analyzing fundamental characteristics of macromolecules in solution, including size and shape, assembly state, and degree of polydispersity. It allows the study of conformational changes and complex formation in physiologically relevant buffers. AUC is useful as a method to characterize structural aspects of macromolecules and as a quantitative technique that provides access to detailed thermodynamic characterization of a system under study. This webinar will describe basic principles underlying sedimentation velocity and sedimentation equilibrium AUC methods and ways in which AUC can be used to complement crystallographic or other structural biology approaches. Examples will be provided to illustrate AUC approaches for characterization of monodisperse or assembling proteins, simultaneous size-and-shape analysis of multi-species mixtures, and determination of affinity and stoichiometry of macromolecular complexes.

    Learning Objectives:
    • Learn about basic principles underlying sedimentation velocity and sedimentation equilibrium AUC methods and ways in which AUC can be used to complement crystallographic or other structural biology approaches
    • Learn AUC approaches for characterization of monodisperse or assembling proteins, simultaneous size-and-shape analysis of multi-species mixtures, and determination of affinity and stoichiometry of macromolecular complexes

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