Identifying glycation hot-spots in bovine milk proteins during production and storage of skim milk powder

We investigated protein glycation in a complex milk system under controlled conditions representative of real-life consumer products, analysing intermediate and final products from skim powder production, aged samples. combined protein-centric LC-MS(/MS) with peptide-centric multi-protease LC-MS/MS focusing on the six most abundant bovine proteins. This strategy resulted identification glycated proteoforms extent per protein, high sequence coverage, relative occupancy sites. identified new hot-spots additionally to ones already described literature. Primary motif analysis revealed that were preceded N-terminally by stretch rich basic amino acids, followed C-terminally enriched aliphatic hydrophobic acids. Our study considerably extends current understanding glycation, discussing their localisation relation primary sequences higher-order structures.