Binding of nucleotides at the active site modulates the local and global conformation of myosin in muscle fibres

D. Lőrinczy, N. Hartvig, N. Farkas, J. Belágyi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres to study the effect of the binding of nucleotides (ADP and AMP·PNP) to myosin. The thermal unfolding of muscle fibres showed three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP·PNP markedly affected the transitions, implying the strong interaction between AMP·PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced only small changes in transition temperatures. Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP·PNP-state resulted in two populations; 50% of labels was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The conformation of the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state to actin might have different local conformations.

Original languageEnglish
Pages (from-to)351-358
Number of pages8
JournalJournal of Thermal Analysis and Calorimetry
Volume65
Issue number2
DOIs
Publication statusPublished - 2001

Fingerprint

Adenylyl Imidodiphosphate
muscle fibers
myosins
adenosine diphosphate
nucleotides
Myosins
Nucleotides
Administrative data processing
Muscle
Conformations
Adenosine Diphosphate
Actins
Fibers
Labels
fibers
suggestion
electron paramagnetic resonance
Deconvolution
thermal stability
heat measurement

Keywords

  • DSC
  • EPR
  • Local and global conformation of myosin
  • Nucleotide binding

ASJC Scopus subject areas

  • Chemical Engineering(all)

Cite this

@article{c20f3ed0a4b047bba71895581da27517,
title = "Binding of nucleotides at the active site modulates the local and global conformation of myosin in muscle fibres",
abstract = "Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres to study the effect of the binding of nucleotides (ADP and AMP·PNP) to myosin. The thermal unfolding of muscle fibres showed three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP·PNP markedly affected the transitions, implying the strong interaction between AMP·PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced only small changes in transition temperatures. Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP·PNP-state resulted in two populations; 50{\%} of labels was highly ordered with respect to fibre axis, whereas the other 50{\%} of labels was randomly oriented. The conformation of the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state to actin might have different local conformations.",
keywords = "DSC, EPR, Local and global conformation of myosin, Nucleotide binding",
author = "D. Lőrinczy and N. Hartvig and N. Farkas and J. Bel{\'a}gyi",
year = "2001",
doi = "10.1023/A:1012400329707",
language = "English",
volume = "65",
pages = "351--358",
journal = "Journal of Thermal Analysis and Calorimetry",
issn = "1388-6150",
publisher = "Springer Netherlands",
number = "2",

}

TY - JOUR

T1 - Binding of nucleotides at the active site modulates the local and global conformation of myosin in muscle fibres

AU - Lőrinczy, D.

AU - Hartvig, N.

AU - Farkas, N.

AU - Belágyi, J.

PY - 2001

Y1 - 2001

N2 - Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres to study the effect of the binding of nucleotides (ADP and AMP·PNP) to myosin. The thermal unfolding of muscle fibres showed three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP·PNP markedly affected the transitions, implying the strong interaction between AMP·PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced only small changes in transition temperatures. Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP·PNP-state resulted in two populations; 50% of labels was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The conformation of the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state to actin might have different local conformations.

AB - Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres to study the effect of the binding of nucleotides (ADP and AMP·PNP) to myosin. The thermal unfolding of muscle fibres showed three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP·PNP markedly affected the transitions, implying the strong interaction between AMP·PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced only small changes in transition temperatures. Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP·PNP-state resulted in two populations; 50% of labels was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The conformation of the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state to actin might have different local conformations.

KW - DSC

KW - EPR

KW - Local and global conformation of myosin

KW - Nucleotide binding

UR - http://www.scopus.com/inward/record.url?scp=0034822828&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034822828&partnerID=8YFLogxK

U2 - 10.1023/A:1012400329707

DO - 10.1023/A:1012400329707

M3 - Article

AN - SCOPUS:0034822828

VL - 65

SP - 351

EP - 358

JO - Journal of Thermal Analysis and Calorimetry

JF - Journal of Thermal Analysis and Calorimetry

SN - 1388-6150

IS - 2

ER -