Impact of A134 and E218 Amino Acid Residues of Tropomyosin on Its Flexibility and Function.

International Journal of Molecular Sciences
Marina MarchenkoAlexander Matyushenko

Abstract

Tropomyosin (Tpm) is one of the major actin-binding proteins that play a crucial role in the regulation of muscle contraction. The flexibility of the Tpm molecule is believed to be vital for its functioning, although its role and significance are under discussion. We choose two sites of the Tpm molecule that presumably have high flexibility and stabilized them with the A134L or E218L substitutions. Applying differential scanning calorimetry (DSC), molecular dynamics (MD), co-sedimentation, trypsin digestion, and in vitro motility assay, we characterized the properties of Tpm molecules with these substitutions. The A134L mutation prevented proteolysis of Tpm molecule by trypsin, and both substitutions increased the thermal stability of Tpm and its bending stiffness estimated from MD simulation. None of these mutations affected the primary binding of Tpm to F-actin; still, both of them increased the thermal stability of the actin-Tpm complex and maximal sliding velocity of regulated thin filaments in vitro at a saturating Ca2+ concentration. However, the mutations differently affected the Ca2+ sensitivity of the sliding velocity and pulling force produced by myosin heads. The data suggest that both regions of instability are esse...Continue Reading

References

Mar 15, 1977·FEBS Letters·A Gorecka, W Drabikowski
Aug 1, 1967·Biochemistry·T Ooi
Jan 1, 1982·Methods in Enzymology·S S Margossian, S Lowey
Jan 1, 1982·Methods in Enzymology·J D Potter
Jan 1, 1999·Die Naturwissenschaften·J C Rüegg
Sep 21, 2001·Journal of Muscle Research and Cell Motility·S V Perry
Feb 5, 2004·Chembiochem : a European Journal of Chemical Biology·Jody M Mason, Katja M Arndt
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Dec 21, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jerry H BrownCarolyn Cohen
Jan 9, 2007·Biophysical Journal·G I Mashanov, J E Molloy
Feb 7, 2007·Advances in Experimental Medicine and Biology·Yasushi NitanaiYuichiro Maéda
Jan 1, 2008·The Journal of Biological Chemistry·John P SumidaSherwin S Lehrer
Jan 16, 2008·Physiological Reviews·Peter GunningEdna Hardeman
Feb 13, 2009·Advances in Experimental Medicine and Biology·E Michael Ostap
Dec 29, 2009·Journal of Structural Biology·Sarah E Hitchcock-DeGregori, Abhishek Singh
Feb 2, 2010·Journal of Structural Biology·Xiaochuan Edward LiStefan Fischer
Apr 22, 2010·Proteins·Kresten Lindorff-LarsenDavid E Shaw
Feb 16, 2011·Biophysical Journal·Roberto Dominguez
Apr 2, 2011·The Journal of Biological Chemistry·Ilya A NevzorovDmitrii I Levitsky
May 19, 2011·Journal of Muscle Research and Cell Motility·Sherwin S LehrerFranklin Fuchs
Jul 27, 2011·Journal of Muscle Research and Cell Motility·Sherwin S LehrerFranklin Fuchs
Feb 22, 2012·Biochemistry. Biokhimii︠a︡·I A Nevzorov, D I Levitsky
Jul 24, 2012·Cell·Elmar BehrmannStefan Raunser
Feb 11, 2014·Journal of Muscle Research and Cell Motility·Mohammed El-Mezgueldi
Jul 22, 2015·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Takeyuki Wakabayashi
Aug 5, 2015·Journal of Cell Science·Peter W GunningDaniel P Mulvihill
Sep 1, 2015·International Review of Cell and Molecular Biology·Sofia Yu Khaitlina
Sep 30, 2015·Annals of Neurology·S DonkervoortC G Bönnemann
Apr 10, 2016·Traffic·Dietmar J Manstein, Daniel P Mulvihill
Apr 12, 2016·Comprehensive Physiology·William Lehman
Jan 20, 2017·Sub-cellular Biochemistry·Sarah E Hitchcock-DeGregori, Bipasha Barua
May 26, 2017·Journal of Muscle Research and Cell Motility·Daniil V ShchepkinAndrey K Tsaturyan
Jun 14, 2017·Proceedings of the National Academy of Sciences of the United States of America·Cristina RisiVitold E Galkin
Dec 27, 2017·The FEBS Journal·Alexander M MatyushenkoDmitrii I Levitsky
Sep 10, 2018·Biophysical Journal·William LehmanMichael J Rynkiewicz
Sep 22, 2018·International Journal of Biological Macromolecules·Alexander M MatyushenkoAndrey K Tsaturyan
Dec 14, 2018·Biochemical and Biophysical Research Communications·Alexander M MatyushenkoDmitrii I Levitsky
Feb 17, 2019·Journal of Muscle Research and Cell Motility·William LehmanJeffrey R Moore
May 11, 2019·Biochemical and Biophysical Research Communications·Victoria V NefedovaAlexander M Matyushenko

❮ Previous
Next ❯

Methods Mentioned

BETA
differential scanning calorimetry
Assay
light scattering
motility assay
PCR
electrophoresis

Software Mentioned

GMimPro
GROMACS
Origin
ImageJ
Tpm
Maxchelator

Related Concepts

Related Feeds

Actin-binding Proteins

Actin-binding proteins are a component of the actin cytoskeleton that play essential roles in cellular functions such as regulation of actin polymerization, maintenance of cell polarity, gene expression regulation, cell motility and many more functions. Discover the latest research on actin-binding proteins here.