Binding of Levosimendan, a Calcium Sensitizer, to Cardiac Troponin C

Research output: Contribution to journalArticle

Abstract

Levosimendan is an inodilatory drug that mediates its cardiac effect by the calcium sensitization of contractile proteins. The target protein of levosimendan is cardiac troponin C (cTnC). In the current work, we have studied the interaction of levosimendan with Ca2+-saturated cTnC by heteronuclear NMR and small angle x-ray scattering. A specific interaction between levosimendan and the Ca2+-loaded regulatory domain of recombinant cTnCC35S was observed. The changes in the NMR spectra of the N-domain of full-length cTnCC35S, due to the binding of levosimendan to the primary site, were indicative of a slow conformational exchange. In contrast, no binding of levosimendan to the regulatory domain of cTnCA-Cys, where all the cysteine residues are mutated to serine, was detected. Moreover, it was shown that levosimendan was in fast exchange on the NMR time scale with a secondary binding site in the C-domain of both cTnCC35S and cTnCA-Cys. The small angle x-ray scattering experiments confirm the binding of levosimendan to Ca2+-saturated cTnC but show no domain-domain closure. The experiments were run in the absence of the reducing agent dithiothreitol and the preservative sodium azide (NaN 3), since we found that levosimendan reacts with these chemicals, commonly used for preparation of NMR protein samples.

Details

Authors
  • Tia Sorsa
  • Sami Heikkinen
  • M. Bret Abbott
  • Ekram Abusamhadneh
  • Tero Laakso
  • Carola Tilgmann
  • Ritva Serimaa
  • Arto Annila
  • Paul R. Rosevear
  • Torbjörn Drakenberg
  • Piero Pollesello
  • Ilkka Kilpeläinen
Organisations
External organisations
  • University of Helsinki
  • University of Cincinnati
  • Orion Corporation
  • VTT Technical Research Centre of Finland, Helsinki
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology
  • Pharmaceutical Chemistry

Keywords

  • Calcium/metabolism*, Magnetic Resonance Spectroscopy, Troponin C/metabolism*, simendan, Myocardium/metabolism*
Original languageEnglish
Pages (from-to)9337-9343
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number12
Publication statusPublished - 2001 Mar 23
Publication categoryResearch
Peer-reviewedYes