Increased Ca2+-sensitivity of the contractile apparatus in end-stage human heart failure results from altered phosphorylation of contractile proteins

Cardiovasc Res. 2003 Jan;57(1):37-47. doi: 10.1016/s0008-6363(02)00606-5.


Objective: The alterations in contractile proteins underlying enhanced Ca(2+)-sensitivity of the contractile apparatus in end-stage failing human myocardium are still not resolved. In the present study an attempt was made to reveal to what extent protein alterations contribute to the increased Ca(2+)-responsiveness in human heart failure.

Methods: Isometric force and its Ca(2+)-sensitivity were studied in single left ventricular myocytes from non-failing donor (n=6) and end-stage failing (n=10) hearts. To elucidate which protein alterations contribute to the increased Ca(2+)-responsiveness isoform composition and phosphorylation status of contractile proteins were analysed by one- and two-dimensional gel electrophoresis and Western immunoblotting.

Results: Maximal tension did not differ between myocytes obtained from donor and failing hearts, while Ca(2+)-sensitivity of the contractile apparatus (pCa(50)) was significantly higher in failing myocardium (deltapCa(50)=0.17). Protein analysis indicated that neither re-expression of atrial light chain 1 and fetal troponin T (TnT) nor degradation of myosin light chains and troponin I (TnI) are responsible for the observed increase in Ca(2+)-responsiveness. An inverse correlation was found between pCa(50) and percentage of phosphorylated myosin light chain 2 (MLC-2), while phosphorylation of MLC-1 and TnT did not differ between donor and failing hearts. Incubation of myocytes with protein kinase A decreased Ca(2+)-sensitivity to a larger extent in failing (deltapCa(50)=0.20) than in donor (deltapCa(50)=0.03) myocytes, abolishing the difference in Ca(2+)-responsiveness. An increased percentage of dephosphorylated TnI was found in failing hearts, which significantly correlated with the enhanced Ca(2+)-responsiveness.

Conclusions: The increased Ca(2+)-responsiveness of the contractile apparatus in end-stage failing human hearts cannot be explained by a shift in contractile protein isoforms, but results from the complex interplay between changes in the phosphorylation status of MLC-2 and TnI.

Publication types

  • Comment
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Atrial Myosins / metabolism
  • Blotting, Western
  • Calcium / metabolism*
  • Cardiac Myosins / metabolism
  • Case-Control Studies
  • Cells, Cultured
  • Contractile Proteins / metabolism*
  • Electrophoresis, Gel, Two-Dimensional
  • Female
  • Heart Failure / metabolism*
  • Humans
  • Male
  • Middle Aged
  • Myocytes, Cardiac / metabolism*
  • Myosin Light Chains / metabolism
  • Phosphorylation
  • Protein Isoforms / metabolism
  • Troponin I / metabolism
  • Troponin T / metabolism


  • Contractile Proteins
  • Myosin Light Chains
  • Protein Isoforms
  • Troponin I
  • Troponin T
  • myosin light chain 2
  • myosin light chain I
  • Atrial Myosins
  • Cardiac Myosins
  • Calcium