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

Strosova, M. and Karlovska, J. and Spickett, C.M. and Grune, T. and Orszagova, Z. and Horakova, L. (2009) Oxidative injury induced by hypochlorous acid to C-ATPase from sarcoplasmic reticulum of skeletal muscle and protective effect of trolox. General Physiology and Biophysics, 28 (2). pp. 195-209. ISSN 0231-5882

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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.

Item type: Article
ID code: 16939
Keywords: Ca-ATPase, HOCl oxidative stress, Trolox, Science (General)
Subjects: Science > Science (General)
Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Related URLs:
    Depositing user: Strathprints Administrator
    Date Deposited: 22 Apr 2010 14:38
    Last modified: 12 Mar 2012 11:06
    URI: http://strathprints.strath.ac.uk/id/eprint/16939

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