Interaction forces between F-actin and titin PEVK domain measured with optical tweezers

Pasquale Bianco, A. Nagy, András Kengyel, Dávid Szatmári, Zsolt Mártonfalvi, Tamás Huber, M. Kellermayer

Research output: Contribution to journalArticle

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Abstract

Titin is a giant protein that determines the elasticity of striated muscle and is thought to play important roles in numerous regulatory processes. Previous studies have shown that titin's PEVK domain interacts with F-actin, thereby creating viscous forces of unknown magnitude that may modulate muscle contraction. Here we measured, with optical tweezers, the forces necessary to dissociate F-actin from individual molecules of recombinant PEVK fragments rich either in polyE or PPAK motifs. Rupture forces at a stretch rate of 250 nm/s displayed a wide, nonnormal distribution with a peak at ∼8 pN in the case of both fragments. Dynamic force spectroscopy experiments revealed low spontaneous off-rates that were increased even by low forces. The loading-rate dependence of rupture force was biphasic for polyE in contrast with the monophasic response observed for PPAK. Analysis of the molecular lengths at which rupture occurred indicated that there are numerous actin-binding regions along the PEVK fragments' contour, suggesting that the PEVK domain is a promiscuous actin-binding partner. The complexity of PEVK-actin interaction points to an adaptable viscoelastic mechanism that safeguards sarcomeric structural integrity in the relaxed state and modulates thixotropic behavior during contraction.

Original languageEnglish
Pages (from-to)2102-2109
Number of pages8
JournalBiophysical Journal
Volume93
Issue number6
DOIs
Publication statusPublished - Sep 2007

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Optical Tweezers
Connectin
Actins
Rupture
Striated Muscle
Elasticity
Muscle Contraction
Spectrum Analysis
Proteins

ASJC Scopus subject areas

  • Biophysics

Cite this

Interaction forces between F-actin and titin PEVK domain measured with optical tweezers. / Bianco, Pasquale; Nagy, A.; Kengyel, András; Szatmári, Dávid; Mártonfalvi, Zsolt; Huber, Tamás; Kellermayer, M.

In: Biophysical Journal, Vol. 93, No. 6, 09.2007, p. 2102-2109.

Research output: Contribution to journalArticle

Bianco, Pasquale ; Nagy, A. ; Kengyel, András ; Szatmári, Dávid ; Mártonfalvi, Zsolt ; Huber, Tamás ; Kellermayer, M. / Interaction forces between F-actin and titin PEVK domain measured with optical tweezers. In: Biophysical Journal. 2007 ; Vol. 93, No. 6. pp. 2102-2109.
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