Reaction of the Hydrated Electron with Histone H1 and Related Compounds Studied by e.s.r. and Spin-trapping

Summary

The reactions of the hydrated electron with histone H1, protamine and related compounds (poly-l-lysine, poly-l-arginine and poly-dl-alanine) were investigated by the spin-trapping technique. In order to identify the radical structure of the spin-adducts originating from macromolecules, the usual spin-trapping technique was developed as follows: N₂-saturated aqueous solutions of proteins containing sodium formate were X-irradiated (4·5 kGy) in the presence of 2-methyl-2-nitrosopropane (MNP) as a spin-trap. The side-products due to the self trapping of MNP radicals were then removed from the spin-adducts of the proteins by a Sephadex G-25 column. Finally the spin-adducts were enzymatically digested to transform the broad e.s.r. signals due to slow tumbling of nitroxyl radicals to identifiable ones. The e.s.r. spectra obtained for all samples showed that the deaminated radical, R—CH—CO—NH— (R:amino acid side chain), was produced.

Furthermore, polyacrylamide gel electrophoresis of the irradiated protamine and histone H1 indicated reduction of molecular size. These results confirm that hydrated electrons react with proteins and induce the deamination reaction which leads to main-chain scission.

• e.s.r.
• spin-trapping
• hydrated electron
• histone H1