Enrichment and identification of 4-HNE modified protein sites by magnetic nanoparticles

• 1、College of Chemistry, Chemical Engineering and Biotechnology, Donghua University
• 2、School of Medicine, Shanghai Jiaotong University, Shanghai, 200025

Abstract：.4-hydroxynonenal is the most abundant and toxic end-products of Lipid peroxidation caused by oxidation damage, the protein adducts of which may cause inflammation, cell death and so on. Identification of 4-HNE protein modified site provides confidential evidence to explain the function of the modification on the protein and its biological effects. However, the enrichment method still need to be improved For this purpose, the present paper has developed hydrazine modified nanoparticles to capture 4-HNE modified peptides from complex samples. Magnetic nanoparticlesFe3O4@SiO2@PSD were designed and synthesized with core-shell structure and PSD as outer layer. Hydrazine were modified in the surface by linker. By reversible chemical reactions between aldehyde and hydrazine under solutions ofdifferent pH, the 4-HNE modified peptides were enriched by the magnetic nanoparticles through the sequent procedure of "reaction, capture and release"on DynaMag Magnet.The method was validated by peptide standards and mixtures of tryptic peptides. It has the potential to identify 4-HNE modification sites in complex samples.

Keywords： 4-HNE modification；Magnetic nanoparticles；LC-MS/MS analysis