Conformational dynamics of titin PEVK explored with FRET spectroscopy.

Abstract:

:The proline-, glutamate-, valine-, and lysine-rich (PEVK) domain of the giant muscle protein titin is thought to be an intrinsically unstructured random-coil segment. Various observations suggest, however, that the domain may not be completely devoid of internal interactions and structural features. To test the validity of random polymer models for PEVK, we determined the mean end-to-end distances of an 11- and a 21-residue synthetic PEVK peptide, calculated from the efficiency of the fluorescence resonance energy transfer (FRET) between an N-terminal intrinsic tryptophan donor and a synthetically added C-terminal IAEDANS acceptor obtained in steady-state and time-resolved experiments. We find that the contour-length scaling of mean end-to-end distance deviates from predictions of a purely statistical polymer chain. Furthermore, the addition of guanidine hydrochloride decreased, whereas the addition of salt increased the FRET efficiency, pointing at the disruption of structure-stabilizing interactions. Increasing temperature between 10 and 50°C increased the normalized FRET efficiency in both peptides but with different trajectories, indicating that their elasticity and conformational stability are different. Simulations suggest that whereas the short PEVK peptide displays an overall random structure, the long PEVK peptide retains residual, loose helical configurations. Transitions in the local structure and dynamics of the PEVK domain may play a role in the modulation of passive muscle mechanics.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Huber T,Grama L,Hetényi C,Schay G,Fülöp L,Penke B,Kellermayer MS

doi

10.1016/j.bpj.2012.08.042

subject

Has Abstract

pub_date

2012-10-03 00:00:00

pages

1480-9

issue

7

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(12)00968-X

journal_volume

103

pub_type

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