Lysozyme fragmentation induced by gamma-radiolysis

Irradiation of lysozyme in frozen states in the absence of oxygen induces specific fragmentation at defined sites along the backbone chain. This paper localizes radio-fragmentation sites by two methods. First, N-terminal sequencing of radiolysis fragments after separation by SDS-polyacrylamide gel electrophoresis and estimation of their molecular masses. Secondly, after purification of radiolysis fragments by reverse phase-HPLC and determination of their molecular mass by electro-spray-ionization mass-spectrometric analysis, combined to N-terminal sequencing and total amino acid analysis. Evidence for the breakage of the peptide bond itself (CO-NH) is given, with radio-fragmentation sites mostly found at the surface of irradiated lysozyme in solvent exposed loops and turns.