Effect of nitric oxide on conformational changes of ovalbumin accompanying self-assembly into non-disease-associated fibrils

Abstract

Ovalbumin (OVA), one of the members of the serpin-superfamily, is the major protein in chicken eggs. Many studies have demonstrated the polymerization ability of OVA but the detailed molecular mechanisms demonstrating its conformational changes accompanying fibril formation are still unclear. This study revealed nitric oxide (NO) induced conformational changes and oligomerization of egg white OVA, resulting in polymerized fibrils. Electron microscopic analysis showed that NO treatment to OVA under mild acidic condition resulted in morphological changes, producing structures similar to the long protein fibrils found in egg white. Spectroscopic analysis and mass spectrometry found that NO-treated OVA contains increased number of β-sheet, indicating transition from α-helixes to β-sheets, and S-nitrosylation of OVA cysteine residue 367. Structural modeling showed that S-nitrosocysteine, Cys367NO, is located in the amyloidogenic core region of the C-terminal region, nearby the N-terminal core region where the α-to-β transition is induced. Such results provide a potential mechanism for non-disease-associated fibril formation of OVA.