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MAY 28, 2015 8:00 AM PDT

Webinar: Quantitative determination of reaction stoichiometry, interaction energies, and solute masses using analytical ultracentrifugation

Speaker
  • Adjunct Faculty Member, Biophysics department at the Virginia Commonwealth University Medical School
    Biography

      Dr. Burgner’s first experience with analytical ultracentrifugation was in 1963 in the laboratory of Frank Gurd at Indiana University Medical School. Dr. Burgner used a Spinco Model E with glass photographic plates and a micro comparator to measure changes in the Schlieren patterns of sperm whale myoglobin in a sedimentation equilibrium experiment.  Dr. Burgner’s next experience occurred some years later in the laboratory of David Filmer at Purdue University around 1975. In this experiment, sedimentation equilibrium was used to determine the stoichiometry and mass of dogfish shark lactate dehydrogenase.  Dr. Burgner earned a Ph.D at Purdue University in the laboratory of William J. Ray Jr. for studies on the reaction mechanisms of lactate dehydrogenase.

      In 1997 Purdue leased an XLA from Beckman Coulter for the purpose of obtaining data for an NIH shared instrumentation grant, and asked Dr. Burgner to oversee the use of the instrument and the grant application.  Approximately two years later, he was awarded the funds to purchase an XLI. He kept the XLA and maintained a laboratory within the structural biology group at Purdue University until late 2005, when a new bioanalysis laboratory was developed in the Bindley Biosciences Center. Dr. Burgner led this facility until retiring from Purdue in 2009. In retirement he volunteered his help to the Biophysics department at the Virginia Commonwealth University Medical School, which had just received a new XLI. Dr. Burgner has continued there as an adjunct faculty member; mostly working on systems involving the interactions of viral proteins with viral proteins, mammalian proteins, and short polymers of DNA and RNA. In addition to these academic experiences, Dr. Burgner taught in Peter Schuck’s course on sedimentation Sedfit at NIH and at the University College London for several years.  He also co-teaches the Proteome Lab XL-A/XL-I Customer Training Course, which now includes basic training on data analysis with SEDFIT.


    Abstract
    With the development of the XLA/XLI analytical ultracentrifuges and their exquisitely sensitive optics that can accurately detect absorbance signals in the range of 0.005 A to 1.5 A. and differences in Rayleigh interference patterns to at least 5 ug/ml of protein solute in solution, new methods for sedimentation data analysis have been developed making it possible to quantitatively determine the stoichiometry of reactions and their energies of interaction as well as solute masses. In this talk, we will consider some of the experimental approaches that Dr. John Burgner and others have used to characterize these processes over the past few years.

    Learning Objectives:
    • Learn new methods for sedimentation data analysis that have been developed making it possible to quantitatively determine the stoichiometry of reactions and their energies of interaction as well as solute masses
    • Learn some of the experimental approaches used to characterize stoichiometry of reactions over the past few years

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