Increased phosphorylation of tropomyosin, troponin I, and myosin light chain-2 after stretch in rabbit ventricular myocardium under physiological conditions.

Abstract

After a change in muscle length, there is an immediate intrinsic response in the amount of developed force, followed by a slower response. Although it has been well documented that the slow force response is at least in part generated by modification of calcium handling, it is unclear whether regulation at the myofilament level occurs during the slow force response. We set out to investigate myofilament calcium sensitivity and phosphorylation status of myofilament proteins after a step-wise change in cardiac muscle length. Ultra-thin right ventricular intact trabeculae were isolated from New Zealand White rabbit hearts and iontophoretically loaded with the calcium indicator bis-fura-2. Twitch force-calcium relationships and steady-state force-[Ca(2+)](i) relationships were measured at various muscle lengths at 37 degrees C using potassium induced contractures. The EC(50) significantly decreased with increase in muscle length and maximal active force development significantly increased, while no significant change in the myofilament cooperativity coefficient was found. Phosphoprotein analysis Pro-Q diamond staining as well as phosphorylation-specific antibodies revealed increased phosphorylation of tropomyosin, troponin I, and myosin light chain-2 at longer muscle lengths. Specifically, TnI phosphorylation at Ser(22/23) was increased. Since the immediate response is seen virtually instantaneously and post-translational modifications are thought not to occur within such a very short timeframe, we hypothesize that these increases in phosphorylation occur during the slow response.

• Authors: Michelle M Monasky, Brandon J Biesiadecki, Paul M L Janssen
• Journal: Journal of molecular and cellular cardiology (J Mol Cell Cardiol) Vol. 48 Issue 5 Pg. 1023-8 (May 2010) ISSN: 1095-8584 [Electronic] England
• PMID: 20298699 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Chemical References

• Myosin Light Chains
• Tropomyosin
• Troponin I
• myosin light chain 2
• Cardiac Myosins
• Calcium

Topics

• Actin Cytoskeleton (metabolism)
• Animals
• Calcium (metabolism)
• Cardiac Myosins (metabolism)
• Electrophoresis, Polyacrylamide Gel
• Heart Ventricles (cytology)
• Male
• Myocardium (metabolism)
• Myosin Light Chains (metabolism)
• Phosphorylation
• Rabbits
• Tropomyosin (metabolism)
• Troponin I (metabolism)