title: Preparation of sequence-controlled triblock copolymer-grafted silica microparticles by sequential-ATRP for highly efficient glycopeptides enrichment.
abstract: As one of the most important subproteomes in eukaryote cells, N-glycoproteins play crucial roles in various of biological processes and have long been considered closely correlated with the occurrence, progression, and metastasis of cancer. Comprehensive characterization of protein N-glycosylation and association of their aberrant patterns to the corresponding cancer stage may provide a unique way to discover new diagnostic biomarkers and therapeutic drug targets. However, the extremely complex nature of biological samples and relatively low abundance of N-glycosylated proteins makes the enrichment of glycoprotein/glycopeptide a prerequisite for large scale N-glycosylation identification. In this work, we prepared sequence controlled triblock copolymer grafted silica-microparticles (TCP-SMs) by sequential atom transfer radical polymerization (sequential-ATRP) of monosaccharides and zwitterionic-ion monomers for highly efficient and selective glycopeptides enrichment. The triblock copolymer is composed of sequence defined poly zwitterionic-ion, poly-N-acetylglucosamine and poly mannose blocks. The glycopolymer blocks carrying densely packed pendent sugars are excellent mimics of the natural carbohydrate clusters and may induce multivalent carbohydrate-carbohydrate interaction (CCI) with the target glycopeptides. Therefore, increased retention of glycopeptides can be expected by the combination of CCI and zwitterionic-HILIC interaction. As a result, 1244 glycopeptides were identified after TCP-SMs enrichment from mouse liver, which are 65-120% higher than that obtained by homoglycopolymer or random-copolymer grafted silica microparticles prepared using the conventional free radical polymerization. These results demonstrate the critical role of sequence-defined block copolymer of TCP-SMs for obtaining enhanced affinity toward glycopeptides and the potential of this sequential-ATRP strategy to integrate different affinity moieties into one enrichment material to achieve deeper coverage in protein PTM mapping.
authors.affiliation: National Center for Protein Sciences Beijing, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine , Beijing 100850, China.
authors.forename: Yiting
authors.initials: Y
authors.lastname: Pan
authors.forename: Cheng
authors.initials: C
authors.lastname: Ma
authors.forename: Wei
authors.initials: W
authors.lastname: Tong
authors.forename: Chao
authors.initials: C
authors.lastname: Fan
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Wanjun
authors.initials: W
authors.lastname: Zhang
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Bo
authors.initials: B
authors.lastname: Peng
authors.forename: Weijie
authors.initials: W
authors.lastname: Qin
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qian
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification*; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification; D006031:Glycosylation; D008099:Liver / Q000378:metabolism*; D051379:Mice; D010316:Particle Size; D058105:Polymerization; D011108:Polymers / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006020:Glycopeptides; D006023:Glycoproteins; D011108:Polymers; D012822:Silicon Dioxide