Analyzing the binding of Co(II)-specific inhibitors to the methionyl aminopeptidases from Escherichia coli and Pyrococcus furiosus

  • Sanghamitra Mitra
  • George Sheppard
  • Jieyi Wang
  • Brian Bennett
  • Richard C. Holz
Original Paper

Abstract

Methionine aminopeptidases (MetAPs) represent a unique class of protease that is capable of the hydrolytic removal of an N-terminal methionine residue from nascent polypeptide chains. MetAPs are physiologically important enzymes; hence, there is considerable interest in developing inhibitors that can be used as antiangiogenic and antimicrobial agents. A detailed kinetic and spectroscopic study has been performed to probe the binding of a triazole-based inhibitor and a bestatin-based inhibitor to both Mn(II)- and Co(II)-loaded type-I (Escherichia coli) and type-II (Pyrococcus furiosus) MetAPs. Both inhibitors were found to be moderate competitive inhibitors. The triazole-type inhibitor was found to interact with both active-site metal ions, while the bestatin-type inhibitor was capable of switching its mode of binding depending on the metal in the active site and the type of MetAP enzyme.

Keywords

Hydrolysis Manganese Electron paramagnetic resonance Antibiotics Electronic absorption 

Notes

Acknowledgments

This work was supported by the National Science Foundation (CHE-0652981, R.C.H.) and the National Institutes of Health (AI056231, B.B.). The Bruker Elexsys spectrometer was purchased by the Medical College of Wisconsin and is supported with funds from the National Institutes of Health (EB001980, B.B.).

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Copyright information

© SBIC 2009

Authors and Affiliations

  • Sanghamitra Mitra
    • 1
    • 4
  • George Sheppard
    • 3
  • Jieyi Wang
    • 3
  • Brian Bennett
    • 2
  • Richard C. Holz
    • 1
  1. 1.Department of ChemistryLoyola University-ChicagoChicagoUSA
  2. 2.Department of Biophysics, The National Biomedical EPR CenterMedical College of WisconsinMilwaukeeUSA
  3. 3.Cancer Research, Global Pharmaceutical R&DAbbott LaboratoriesAbbott ParkUSA
  4. 4.Department of ChemistryBoston UniversityBostonUSA

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