Cardiac troponin T forms a tetramer in vitro

Karim C. Lounes, Borries Demeler, David E. Anderson, Aldrin V Gomes, James D. Potter, Rashid Nassar, Page A W Anderson

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Cardiac troponin T (cTnT) is a myofibrillar protein essential for calcium-dependent contraction. This property has led to functional studies of developmentally expressed cTnT isoforms and mutants identified in patients with hypertrophic cardiomyopathy. The release of cTnT into the serum following myocardial infarction has led to the development of antibody-based assays for measuring cTnT serum concentration. We examined the behavior of cTnT in solution. Recombinant human cTnT3, the dominant isoform in the adult human heart, was used. The protein was pure and functional, as demonstrated by SDS-PAGE and surface plasmon resonance. cTnT3 was found to bind specifically and in a concentration-dependent manner to cTnC. Routine size exclusion chromatography suggested a higher-than-expected MW for cTnT. Using analytical ultracentrifugation, we found cTnT3 in solution to be mainly in the form of a tightly bound tetramer at concentrations as low as 4 μmol/L. Our sedimentation velocity and transmission electron microscopy results indicate that the tetramer's shape is elongated rather than globular. CTnT's self-association in solution is an important consideration in the design and interpretation of experiments with the aim of understanding the biochemical and biophysical properties of cTnT, its isoforms, and its mutants.

Original languageEnglish (US)
Pages (from-to)1970-1976
Number of pages7
JournalBiochemistry
Volume47
Issue number7
DOIs
StatePublished - Feb 19 2008

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Troponin T
Protein Isoforms
Surface Plasmon Resonance
Size exclusion chromatography
Hypertrophic Cardiomyopathy
Ultracentrifugation
Surface plasmon resonance
In Vitro Techniques
Serum
Transmission Electron Microscopy
Sedimentation
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Assays
Proteins
Myocardial Infarction
Association reactions
Transmission electron microscopy
Calcium
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lounes, K. C., Demeler, B., Anderson, D. E., Gomes, A. V., Potter, J. D., Nassar, R., & Anderson, P. A. W. (2008). Cardiac troponin T forms a tetramer in vitro. Biochemistry, 47(7), 1970-1976. https://doi.org/10.1021/bi7012596

Cardiac troponin T forms a tetramer in vitro. / Lounes, Karim C.; Demeler, Borries; Anderson, David E.; Gomes, Aldrin V; Potter, James D.; Nassar, Rashid; Anderson, Page A W.

In: Biochemistry, Vol. 47, No. 7, 19.02.2008, p. 1970-1976.

Research output: Contribution to journalArticle

Lounes, KC, Demeler, B, Anderson, DE, Gomes, AV, Potter, JD, Nassar, R & Anderson, PAW 2008, 'Cardiac troponin T forms a tetramer in vitro', Biochemistry, vol. 47, no. 7, pp. 1970-1976. https://doi.org/10.1021/bi7012596
Lounes KC, Demeler B, Anderson DE, Gomes AV, Potter JD, Nassar R et al. Cardiac troponin T forms a tetramer in vitro. Biochemistry. 2008 Feb 19;47(7):1970-1976. https://doi.org/10.1021/bi7012596
Lounes, Karim C. ; Demeler, Borries ; Anderson, David E. ; Gomes, Aldrin V ; Potter, James D. ; Nassar, Rashid ; Anderson, Page A W. / Cardiac troponin T forms a tetramer in vitro. In: Biochemistry. 2008 ; Vol. 47, No. 7. pp. 1970-1976.
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