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Muscle Biophysics pp 163–174Cite as

Extraction and Replacement of the Tropomyosin–Troponin Complex in Isolated Myofibrils

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 682))

Abstract

Tropomyosin (Tm) is an essential component in the regulation of striated muscle contraction. Questions about Tm functional role have been difficult to study because sarcomere Tm content is not as easily manipulated as Troponin (Tn). Here we describe the method we recently developed to replace Tm-Tn of skeletal and cardiac myofibrils from animals and humans to generate an experimental model of homogeneous Tm composition and giving the possibility to measure a wide range of mechanical parameters of contraction (e.g. maximal force and kinetics of force generation). The success of the exchange was determined by SDS–PAGE and by mechanical measurements of calcium dependent force activation on the reconstituted myofibrils. In skeletal and cardiac myofibrils, the percentage of Tm replacement was higher than 90%. Maximal isometric tension was 30–35% lower in the reconstituted myofibrils than in control myofibrils but the rate of force activation (kACT) and that of force redevelopment (kTR) were not significantly changed. Preliminary results show the effectiveness of Tm replacement in human cardiac myofibrils. This approach can be used to test the functional impact of Tm mutations responsible for human cardiomyopathies.

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Acknowledgement

This work was supported by 7th Framework Programs of the European Union (STREP Project “BIG-HEART”, grant agreement 241577) and by Telethon-Italy (GGP07133). Authors gratefully acknowledge Dr. Earl Homsher for helpful discussion and technical advice.

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Authors and Affiliations

  1. Dipartimento di Scienze Fisiologiche, Università di Firenze, Viale Morgagni 63, Firenze, I-50134, Italy

    Chiara Tesi

Authors
  1. Beatrice Scellini
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  2. Nicoletta Piroddi
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  3. Corrado Poggesi
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  4. Chiara Tesi
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Corresponding author

Correspondence to Chiara Tesi .

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Editors and Affiliations

  1. McGill University, Pine Avenue West 475, Montreal, H2W 1S4, Québec, Canada

    Dilson E. Rassier

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Scellini, B., Piroddi, N., Poggesi, C., Tesi, C. (2010). Extraction and Replacement of the Tropomyosin–Troponin Complex in Isolated Myofibrils. In: Rassier, D. (eds) Muscle Biophysics. Advances in Experimental Medicine and Biology, vol 682. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-6366-6_9

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