Investigation of a genetically engineered mutant of barnacle troponin C containing a central helix deletion

L. D. Allhouse, T. Miller, Q. Li, G. Guzman, J. D. Potter, A. Mandveno, C. C. Ashley

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

To examine the importance of the central α-helix of troponin C (TnC) we have bacterially expressed one of the isoforms of barnacle TnC (BTnC2), BTnCWT, but without the aspartate residue at position 80 in the central helix (BTnC80-). This manipulation is expected to produce an approximately 100°axial rotation of the C-domain with respect to the N-domain, and a net charge change of -1. BTnC80- mutant was able to restore force to TnC-depleted skinned barnacle myofibrillar bundles to a greater extent than wild-type protein (≃170%). Competition experiments between BTnC80- and BTnC2-4-, a mutant lacking both of the calcium-specific sites (sites II and IV), shows that deletion of a single amino acid in the central helix results in a protein with increased affinity for the thin filament and one that is bound preferentially compared to BTnC2-4- when at equimolar concentrations.

Original languageEnglish
Pages (from-to)67-75
Number of pages9
JournalPflugers Archiv European Journal of Physiology
Volume439
Issue number1-2
DOIs
StatePublished - Dec 1 1999
Externally publishedYes

Fingerprint

Thoracica
Troponin C
Aspartic Acid
Protein Isoforms
Proteins
Calcium
Amino Acids
Experiments

Keywords

  • Barnacle
  • Central-helix troponin C
  • Contraction
  • Mutagenesis

ASJC Scopus subject areas

  • Physiology

Cite this

Investigation of a genetically engineered mutant of barnacle troponin C containing a central helix deletion. / Allhouse, L. D.; Miller, T.; Li, Q.; Guzman, G.; Potter, J. D.; Mandveno, A.; Ashley, C. C.

In: Pflugers Archiv European Journal of Physiology, Vol. 439, No. 1-2, 01.12.1999, p. 67-75.

Research output: Contribution to journalArticle

Allhouse, L. D. ; Miller, T. ; Li, Q. ; Guzman, G. ; Potter, J. D. ; Mandveno, A. ; Ashley, C. C. / Investigation of a genetically engineered mutant of barnacle troponin C containing a central helix deletion. In: Pflugers Archiv European Journal of Physiology. 1999 ; Vol. 439, No. 1-2. pp. 67-75.
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