Oxidative injury induced by hypochlorous acid to Ca-ATPase from sarcoplasmic reticulum of skeletal muscle and protective effect of trolox

Miriam Strosova, Jana Karlovska, Corinne M. Spickett, Tilman Grune, Zuzana Orszagova, Zuzana Horakova

Research output: Contribution to journalArticle

Abstract

Hypochlorous acid (HOCl) concentration-dependently decreased ATPase activity and SH groups of pure Ca-ATPase from sarcoplasmic reticulum (SERCA) of rabbit skeletal muscle with IC(50) of 150 micromol/l and 6.6 micromol/l, respectively. This indicates that SH groups were not critical for impairment of Ca-ATPase activity. Pure Ca-ATPase activity was analysed individually with respect to both substrates, Ca(2+) and ATP. Concerning dependence of ATPase activity on HOCl (150 micromol/l) as a function of free Ca(2+) and ATP, V(max) of both dependences decreased significantly, while the affinities to individual substrates were not influenced, with the exception of the regulatory binding site of ATP. On increasing HOCl concentration, fluorescence of fluorescein-5-isothiocyanate (FITC) decreased, indicating binding of HOCl to nucleotide binding site of SERCA. A new fragment appeared at 75 kDa after HOCl oxidation of SR, indicating fragmentation of SERCA. Fragmentation may be associated with protein carbonyl formation. The density of protein carbonyl bands at 75 and 110 kDa increased concentration- and time-dependently. Trolox (250 micromol/l) recovered the Ca-ATPase activity decrease induced by HOCl, probably by changing conformational properties of the Ca-ATPase protein. Trolox inhibited FITC binding to SERCA.
LanguageEnglish
Pages195-209
Number of pages15
JournalGeneral Physiology and Biophysics
Volume28
Issue number2
DOIs
Publication statusPublished - Jun 2009

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Hypochlorous Acid
Sarcoplasmic Reticulum
Adenosine Triphosphatases
Skeletal Muscle
Wounds and Injuries
Fluorescein-5-isothiocyanate
Adenosine Triphosphate
Binding Sites
Protein Carbonylation
6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
Proteins
Nucleotides
Fluorescence
Rabbits

Keywords

  • Ca-ATPase
  • HOCl oxidative stress
  • Trolox
  • Science (General)

Cite this

Strosova, Miriam ; Karlovska, Jana ; Spickett, Corinne M. ; Grune, Tilman ; Orszagova, Zuzana ; Horakova, Zuzana. / Oxidative injury induced by hypochlorous acid to Ca-ATPase from sarcoplasmic reticulum of skeletal muscle and protective effect of trolox. In: General Physiology and Biophysics. 2009 ; Vol. 28, No. 2. pp. 195-209.
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Oxidative injury induced by hypochlorous acid to Ca-ATPase from sarcoplasmic reticulum of skeletal muscle and protective effect of trolox. / Strosova, Miriam; Karlovska, Jana; Spickett, Corinne M.; Grune, Tilman; Orszagova, Zuzana; Horakova, Zuzana.

In: General Physiology and Biophysics, Vol. 28, No. 2, 06.2009, p. 195-209.

Research output: Contribution to journalArticle

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