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S-NO-actin: S-nitrosylation kinetics and the effect on isolated vascular smooth muscle

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Abstract

We describe the modification of reactive actin sulfhydryls by S-nitrosoglutathione. Kinetics of S-nitrosylation and denitrosylation suggest that only one cysteine of actin is involved in the reactions. By using the bifunctional sulfhydryl cross-linking reagent N,N′-1,4-phenylenebismaleimide and the monofunctional reagent N-iodoacetyl-N′-(5-sulpho-1-naphthyl)ethylenediamine, we identified this residue as Cys374. The time course of filament formation followed by high-shear viscosity changes revealed that S-nitrosylated G-actin polymerizes less efficiently than native monomers. The observed decrease in specific viscosity at steady state is due mainly to a marked inhibition of filament end-to-end annealing and, partially, to a reduction in F-actin concentration. Finally, S-nitrosylated actin acts as nitric oxide donor showing a fast, potent vasodilating activity at unusually low concentrations, being comparable with that of low molecular weight nitrosothiols.

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Authors and Affiliations

  1. Lab. Biochem. Biophys. Cytoskel., Department of Biology, University of Milan, via Celoria 26, I-20133, Milan, Italy

    Isabella Dalle-donne, Aldo Milzani/snm>  & Roberto Colombo

  2. Pharmacology Section, Institute for Mental and Nervous Diseases, University of Siena, via A. Moro 4, I-53100, Siena, Italy

    Daniela Giustarini, Paolo Di Simplicio & Ranieri Rossi

Authors
  1. Isabella Dalle-donne
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  2. Aldo Milzani/snm>
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  3. Daniela Giustarini
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  4. Paolo Di Simplicio
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  5. Roberto Colombo
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  6. Ranieri Rossi
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Dalle-donne, I., Milzani/snm> , A., Giustarini, D. et al. S-NO-actin: S-nitrosylation kinetics and the effect on isolated vascular smooth muscle. J Muscle Res Cell Motil 21, 171–181 (2000). https://doi.org/10.1023/A:1005671319604

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