Conformational changes of troponin C within the thin filaments detected by neutron scattering

Journal of Molecular Biology
Fumiko MatsumotoSatoru Fujiwara

Abstract

Regulation of skeletal and cardiac muscle contraction is associated with structural changes of the thin filament-based proteins, troponin consisting of three subunits (TnC, TnI, and TnT), tropomyosin, and actin, triggered by Ca2+-binding to TnC. Knowledge of in situ structures of these proteins is indispensable for elucidating the molecular mechanism of this Ca2+-sensitive regulation. Here, the in situ structure of TnC within the thin filaments was investigated with neutron scattering, combined with selective deuteration and the contrast matching technique. Deuterated TnC (dTnC) was first prepared, this dTnC was then reconstituted into the native thin filaments, and finally neutron scattering patterns of these reconstituted thin filaments containing dTnC were measured under the condition where non-deuterated components were rendered "invisible" to neutrons. The obtained scattering curves arising only from dTnC showed distinct difference in the absence and presence of Ca2+. These curves were analyzed by model calculations using the Monte Carlo method, in which inter-dTnC interference was explicitly taken into consideration. The model calculation showed that in situ radius of gyration of TnC was 23 A (99% confidence limits betwee...Continue Reading

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Citations

Jun 2, 2009·The Journal of Biological Chemistry·Tomoyoshi KobayashiMinae Kobayashi
Aug 11, 2010·The Journal of Biological Chemistry·Ahmed HoumeidaHoward D White
Jan 31, 2014·Proceedings of the National Academy of Sciences of the United States of America·Ji Young MunRoger Craig
Mar 12, 2009·Biochemical and Biophysical Research Communications·Fumiko MatsumotoSatoru Fujiwara
Jan 27, 2007·Journal of Molecular Biology·Satoru Fujiwara, Fumiko Matsumoto
Dec 18, 2007·Biochemical and Biophysical Research Communications·Yasunobu SugimotoKatsuzo Wakabayashi
Oct 19, 2005·Advances in Protein Chemistry·Jerry H Brown, Carolyn Cohen
Feb 16, 2005·Journal of Muscle Research and Cell Motility·Monica X LiBrian D Sykes
Sep 4, 2020·Micromachines·Wataru MinoshimaYasushi Inouye

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