DSC and EPR study on AMP.PNP, BeFx and AlF4 containing myosin nucleotide complexes

Research output: Contribution to journalArticle

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Abstract

Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated skeletal muscle fibres to study the effect of the binding of nucleotides and nucleotide analogues to myosin. The thermal unfolding of muscle fibres in rigor showed three discrete domain regions with thermal stability of 52.2, 58.8 and 67.8°C. AMP.PNP and ATP plus AlF3 or BeF2 affected markedly the transitions, which implies the strong interaction between AMP.PNP or nucleotide analogues and catalytic domain of myosin, and a partial dissociation of heads from actin. ADP.BeFx and ADP.AlF4- states model the transition states of the ATP hydrolysis cycle which precede the powerstroke of the muscle fibres. Spectrum deconvolution 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 myosin heads which showed high degree of order were in the strongly binding ADP-state. The spectra in ADP.AlF4- - and ADP.BeFx state reflected random orientation of labels with increased rotational mobility in comparison with rigor. The results suggest that myosin in muscle fibres in ADP.BeFx state exists in two forms.

Original languageEnglish
Pages (from-to)565-572
Number of pages8
JournalJournal of Thermal Analysis and Calorimetry
Volume72
Issue number2
DOIs
Publication statusPublished - 2003

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adenosine monophosphate
myosins
Administrative data processing
adenosine diphosphate
nucleotides
Adenosine Monophosphate
Myosins
Nucleotides
muscle fibers
Paramagnetic resonance
Adenosine Diphosphate
Muscle
Fibers
Labels
adenosine triphosphate
Adenosinetriphosphate
Adenosine Triphosphate
analogs
skeletal muscle
fibers

Keywords

  • Aluminium and beryllium fluoride
  • ATP hydrolysis
  • Conformation of myosin
  • DSC
  • EPR
  • Nucleotide-myosin interaction

ASJC Scopus subject areas

  • Chemical Engineering(all)

Cite this

DSC and EPR study on AMP.PNP, BeFx and AlF4 containing myosin nucleotide complexes. / Lőrinczy, D.; Kiss, M.; Belágyi, J.

In: Journal of Thermal Analysis and Calorimetry, Vol. 72, No. 2, 2003, p. 565-572.

Research output: Contribution to journalArticle

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