Radiolytic and enzymatic dimerization of tyrosyl residues in insulin, ribonuclease, papain and collagen

Int J Radiat Biol Relat Stud Phys Chem Med. 1983 Mar;43(3):249-65. doi: 10.1080/09553008314550301.

Abstract

Insulin, ribonuclease, papain and collagen solutions saturated with nitrogen, N2O or air were irradiated with doses of 10 to 640 Gy of gamma rays. Protein solutions were also oxidized enzymatically in a system of horse-radish peroxidase: hydrogen peroxide. Column chromatography (Sephadex G-75 or Sephacryl S-200) of treated protein solutions revealed that they contain protein molecular aggregates. Nitrogen saturation of solution before irradiation was most favourable for radiation-induced aggregation of proteins. Fluorescence analysis of protein solutions resulted in detection of dityrosyl structures in irradiated as well as in enzymatically oxidized proteins. Concentration of dityrosine in proteins studied was determined fluorimetrically in their hydrolysates separated on BioGel P-2 column. In irradiated proteins, dityrosine was present almost exclusively in their aggregated forms. In proteins oxidized enzymatically, dityrosine was also present in fractions containing apparently unchanged protein. Mechanisms which could account for differences in the yield of dityrosine formation in radiolysis and in enzymatic oxidation of proteins are suggested.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cobalt Radioisotopes
  • Collagen / radiation effects*
  • Gamma Rays
  • Horseradish Peroxidase
  • Hydrogen Peroxide
  • Insulin / radiation effects*
  • Papain / radiation effects*
  • Ribonucleases / radiation effects*
  • Tyrosine / analogs & derivatives*
  • Tyrosine / analysis

Substances

  • Cobalt Radioisotopes
  • Insulin
  • Tyrosine
  • Collagen
  • Hydrogen Peroxide
  • dityrosine
  • Horseradish Peroxidase
  • Ribonucleases
  • Papain