Application of X-ray absorption spectroscopy and anomalous small angle scattering to RNA polymerase

L. Powers, F. Y.H. Wu, J. C. Phillips, W. J. Huang, R. B. Sinclair

Research output: Contribution to journalArticle

Abstract

X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (≤ ∼ 5 Å) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the EXAFS region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near-physiological concentrations (as low as ∼ 100 μM). With careful choice of model compounds, judicious use of fitting procedures, and consideration of the results of biochemical and other spectroscopic results, this data has provided pivotal information about the structures of these active sites which store energy in their conformation changes or ligand exchanges. Although the application of anomalous small angle scattering to biomolecules has occurred more recently, it clearly provides a method of determining distances between active sites that are outside the range of X-ray absorption spectroscopy. The wavelength dependence of the X-ray scattering power varies rapidly near the edge of the absorbing atom in both amplitude and phase. This behavior selectively alters the contribution of the absorbing atom to the scattering pattern. The structure-function relationship of the intermediate states provide the key to understanding the mechanisms of these complex molecules. It is this precise structural information about the active sites that is not obtainable by other spectroscopic techniques. Combination of these techniques offers a unique approach to the determination of the organization of active sites in biomolecules, especially metalloenzymes. Application of these methods to the substrate and template binding sites of RNA polymerase which contain zinc atoms demonstrates the versatility of this approach.

Original languageEnglish (US)
Pages (from-to)36-40
Number of pages5
JournalNuclear Inst. and Methods in Physics Research, A
Volume291
Issue number1-2
DOIs
StatePublished - May 20 1990

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ASJC Scopus subject areas

  • Nuclear and High Energy Physics
  • Instrumentation

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