Mass spectrometric determination of association constants of adenylate kinase with two noncovalent inhibitors

  • Jürg M. Daniel
  • Gregor McCombie
  • Silke Wendt
  • Renato Zenobi
Focus: Noncovalent Interactions

Abstract

Noncovalent complexes between chicken muscle adenylate kinase and two inhibitors, P1,P4-di(adenosine-5′)tetraphosphate (Ap4A) and P1,P5-di(adenosine-5′) pentaphosphate (Ap5A), were investigated with electrospray ionization mass spectrometry under non-denaturing conditions. The nonconvalent nature and the specificity of the complexes are demonstrated with a number of control experiments. Titration experiments allowed the association constants for inhibitor binding to be determined. Problems with concentration dependent ion yields are circumvented by a data evaluation method that is insensitive to the overall ionization efficiency. The Ka values found were 9. 0 × 104 M−1 (Ap4A) and 4. 0 × 107 M−1 (Ap5A), respectively, in very good agreement with available literature data.

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Copyright information

© American Society for Mass Spectrometry 2003

Authors and Affiliations

  • Jürg M. Daniel
    • 1
  • Gregor McCombie
    • 1
  • Silke Wendt
    • 1
  • Renato Zenobi
    • 1
  1. 1.Department of Chemistry, Swiss Federal Institute of TechnologyETHZürichSwitzerland

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