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Fluorescence Spectroscopy, Imaging and Probes pp 277–293Cite as

Emission Spectroscopy of Complex Formation between Escherichia coli Purine Nucleoside Phosphorylase (PNP) and Identified Tautomeric Species of Formycin Inhibitors Resolves Ambiguities Found in Crystallographic Studies

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Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 2))

Abstract

The ubiquitous purine nucleoside phosphorylases (PNPs) play a key role in the purine salvage pathway, and are widely considered as major targets in the chemotherapy of immune system diseases, as well as in potentiation of the antitumor and antiviral activities of therapeutically active nucleoside analogues, by preventing them from phosphorolytic cleavage. Therefore, wide attention is devoted to development of more potent and specific inhibitors of the enzyme from various sources, and to studies of the mechanism of enzyme action. This review recalls the results of studies by steadystate and time-resolved emission (fluorescence and phosphorescence) spectroscopy, X-ray crystallography and enzyme kinetics on the interaction of highly purified bacterial (E. coli) purine nucleoside phosphorylase with a specific formycin A inhibitor (antibiotic) and its N-methylated analogues. The red shift of absorption and emission spectra of the ligands versus the enzyme permits selective excitation of ligand in the enzyme-ligand complex, as well as selective detection of enzyme or ligand emission.

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Authors
  1. B. Kierdaszuk
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Editors and Affiliations

  1. Institute of Molecular Biological Sciences, Amsterdam, Vrije Universiteit, De Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands

    Ruud Kraayenhof

  2. MicroSpectroscopy Center, Wageningen University, Dreijenlaan 3, 6703 HA, Wageningen, The Netherlands

    Antonie J. W. G. Visser

  3. Dept. of Molecular Biophysics, University of Utrecht, Princetonplein 1, 3584 CC, Utrecht, The Netherlands

    Hans C. Gerritsen

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© 2002 Springer-Verlag Berlin Heidelberg

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Kierdaszuk, B. (2002). Emission Spectroscopy of Complex Formation between Escherichia coli Purine Nucleoside Phosphorylase (PNP) and Identified Tautomeric Species of Formycin Inhibitors Resolves Ambiguities Found in Crystallographic Studies. In: Kraayenhof, R., Visser, A.J.W.G., Gerritsen, H.C. (eds) Fluorescence Spectroscopy, Imaging and Probes. Springer Series on Fluorescence, vol 2. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-56067-5_17

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  • DOI: https://doi.org/10.1007/978-3-642-56067-5_17

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-62732-3

  • Online ISBN: 978-3-642-56067-5

  • eBook Packages: Springer Book Archive

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