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Analytical Ultracentrifugation and its use in Biotechnology

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Modern Analytical Ultracentrifugation

Part of the book series: Emerging Biochemical and Biophysical Techniques ((EBBT))

Abstract

Analytical ultracentrifugation has played a critical role in laying the foundations for modern molecular biology. Among its achievements are the demonstration that proteins are macromolecules rather than complexes of smaller units (Svedberg and Fahraeus 1926), and direct support for the semi-conservative replication of DNA as proposed by Watson and Crick (Meselson and Stahl 1958). Unlike techniques such as SDS polyacrylamide gel electrophoresis (SDS PAGE) or gel permeation chromatography (GPC), analytical ultracentrifugation can be used to determine absolute molecular weights without the use of molecular weight standards or interference from the sieving matrix used for separation. Mass spectrometry technology has improved tremendously over the last 5 years and routinely enables researchers to determine molecular weights of macromolecules far more accurately than by analytical ultracentrifugation. However, molecular weights of associating macromolecules in solution are still best determined by centrifugation. Quantitation of these interactions by determining association constants is most easily done by sedimenting solutions to equilibrium and fitting the resulting concentration gradient to a specific association model. The interactions between molecules in oligomeric proteins, self-associating proteins, and interacting systems such as receptor-ligand complexes can be investigated by analytical ultracentrifugation.

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Authors
  1. Steven J. Shire
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Editors and Affiliations

  1. Department of Molecular & Cell Biology and Analytical Ultracentrifugation Facility, University of Connecticut, 75 North Eagleville Road, 06269-3125, Storrs, CT, USA

    Todd M. Schuster

  2. Department of Biochemistry, University of New Hampshire, 03824, Durham, NH, USA

    Thomas M. Laue

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© 1994 Birkhäuser Boston

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Shire, S.J. (1994). Analytical Ultracentrifugation and its use in Biotechnology. In: Schuster, T.M., Laue, T.M. (eds) Modern Analytical Ultracentrifugation. Emerging Biochemical and Biophysical Techniques. Birkhäuser Boston. https://doi.org/10.1007/978-1-4684-6828-1_15

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  • DOI: https://doi.org/10.1007/978-1-4684-6828-1_15

  • Publisher Name: Birkhäuser Boston

  • Print ISBN: 978-1-4684-6830-4

  • Online ISBN: 978-1-4684-6828-1

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