Modification of creatine kinase by S-nitrosothiols: S-nitrosation vs. S-thiolation

Free Radic Biol Med. 2000 Jun 1;28(11):1671-8. doi: 10.1016/s0891-5849(00)00281-1.


Creatine kinase is reversibly inhibited by incubation with S-nitrosothiols. Loss of enzyme activity is associated with the depletion of 5,5'-dithiobis (2-nitrobenzoic acid)-accessible thiol groups, and is not due to nitric oxide release from RSNO. Full enzymatic activity and protein thiol content are restored by incubation of the S-nitrosothiol-modified protein with glutathione. S-nitroso-N-acetylpenicillamine, which contains a more sterically hindered S-nitroso group than S-nitrosoglutathione, predominantly modifies the protein thiol to an S-nitrosothiol via a transnitrosation reaction. In contrast, S-nitrosoglutathione modifies creatine kinase predominantly by S-thiolation. Both S-nitroso-N-acetylpenicillamine and S-nitrosoglutathione modify bovine serum albumin to an S-nitroso derivative. This indicates that S-thiolation and S-nitrosation are both relevant reactions for S-nitrosothiols, and the relative importance of these reactions in biological systems depends on both the environment of the protein thiol and on the chemical nature of the S-nitrosothiol.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cattle
  • Creatine Kinase / antagonists & inhibitors*
  • Creatine Kinase / chemistry*
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology
  • Free Radicals / chemistry
  • Glutathione / analogs & derivatives
  • Glutathione / chemistry
  • Glutathione / pharmacology
  • In Vitro Techniques
  • Nitroso Compounds / chemistry
  • Nitroso Compounds / pharmacology*
  • Penicillamine / analogs & derivatives
  • Penicillamine / chemistry
  • Penicillamine / pharmacology
  • Rabbits
  • S-Nitroso-N-Acetylpenicillamine
  • S-Nitrosoglutathione
  • Serum Albumin, Bovine / chemistry
  • Sulfhydryl Compounds / chemistry
  • Sulfhydryl Compounds / pharmacology*


  • Enzyme Inhibitors
  • Free Radicals
  • Nitroso Compounds
  • Sulfhydryl Compounds
  • Serum Albumin, Bovine
  • S-Nitrosoglutathione
  • S-Nitroso-N-Acetylpenicillamine
  • Creatine Kinase
  • Glutathione
  • Penicillamine