S-Nitrosylation Decreases the Adsorption of H-Ras in Lipid Bilayer and Changes Intrinsic Catalytic Activity

• Published in: Cell biochemistry and biophysics Vol. 59; no. 3; pp. 191 - 199
• Main Authors: Shanshiashvili, L, Narmania, N, Barbakadze, T, Zhuravliova, E, Natsvlishvili, N, Ramsden, J, Mikeladze, D. G
• Format: Journal Article
• Language: English
• Published: New York New York : Humana Press Inc 01-04-2011; Humana Press Inc; Springer Nature B.V
• Subjects: Adsorption; Animals; Biocatalysis; Biochemistry; Biological and Medical Physics; Biomedical and Life Sciences; Biophysics; Biotechnology; Cattle; Cell Biology; Cysteine - analogs & derivatives; Cysteine - chemistry; Cysteine - genetics; Cysteine - metabolism; Farnesylation; GTPase activity; Kinetics; Life Sciences; Lipid absorption; Lipid Bilayers - chemistry; Lipid Bilayers - metabolism; Lipids; Mutation; Original Paper; Oxidation-Reduction; Pharmacology/Toxicology; Ras protein; ras Proteins - chemistry; ras Proteins - genetics; ras Proteins - metabolism; S-Nitrosothiols - chemistry; S-Nitrosothiols - metabolism; S-nitrosylation; Ras protein; GTPase activity; Farnesylation; Lipid absorption; S-nitrosylation
• ISSN: 1085-9195; 1559-0283
• DOI: 10.1007/s12013-010-9132-x

Structural, chemical, and mutational studies have shown that C-terminal cysteine residues on H-Ras could potentially be oxidized by nitrosylation. For investigating the effect of nitrosylation of Ras molecule on the adsorption of farnesylated H-Ras into lipid layer, experiments with optical waveguide lightmode spectroscopy were used. The analysis of association/dissociation kinetics to planar phospholipids under controlled hydrodynamic conditions has shown that preliminary treatment of protein by S-nitroso-cysteine decreased the adsorption of farnesylated H-Ras. The authors have found that compared with nitrosylated forms, farnesylated H-Ras has more compact configuration, because of the smaller area occupied by protein upon absorption at the membrane. The association rate coefficient for unmodified H-Ras was lower than similar parameter for farnesylated and nitrosylated forms. However, the desorbability, i.e., parameter, which reflects the rate of dissociation of protein from lipids is higher for farnesylated H-Ras. In addition, it was have found that farnesylation of cytoplasmic H-Ras, in contrast to membrane-derived forms, inhibits intrinsic GTPase activity of protein, and preliminary treatment of H-Ras by S-nitroso-cysteine restores the activity to the control level. These data suggest that nitrosylation of H-Ras rearranges the adsorptive potential and intrinsic GTPase activity of H-Ras through modification of C-terminal cysteines of molecule.