A promising nanoprobe based on hydrophilic interaction liquid chromatography and immobilized metal affinity chromatography for capture of glycopeptides and phosphopeptides

• Published in: Analytica chimica acta 2019-08, Vol.1067, p.1-10
• Main Authors: Wang, Zidan, Wang, Jiawen, Sun, Nianrong, Deng, Chunhui
• Format: Article
• Language: English
• Subjects: Affinity chromatography; Animals; Antibiotics; Brain; Cataracts; Chromatography; Chromatography, Affinity; Chromatography, Liquid; Cysteine - chemistry; Eye diseases; Eye lens; Glycan; Glycopeptides; Glycopeptides - isolation & purification; Glycoproteomics; Glycosylation; Gold - chemistry; Humans; Hydrophilicity; Hydrophobic and Hydrophilic Interactions; Indoles - chemistry; Ions - chemistry; L-cysteine; Lens, Crystalline - chemistry; Liquid chromatography; Mice; Multi enrichment mechanism; Nanoparticles - chemistry; Peptides; Phosphopeptides - isolation & purification; Phosphoproteomics; Phosphorylation; Polymers - chemistry; Selectivity; Simultaneous capture; Titanium; Titanium - chemistry; Titanium ion
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2019.04.010

Glycosylation and phosphorylation are two important mechanisms in organism, both of which can finely regulate protein function respectively or simultaneously. Functional nanoprobes with various reversible interactions of aiming at glycan or phosphate group have been advanced dramatically, however, the comprehensive analysis towards them is largely restricted by the challenge to capture glycopeptides and phosphopeptides at the same time from complex samples due to the limitation of nanoprobe functionalization. Here, we fabricate a promising nanoprobe using the hydrophilic l-cysteine and titanium ions to enhance the phosphopeptides capture, meanwhile, realize the glycopeptides capture. We test the performance of the promising nanoprobe, showing that it is not only equipped with good unbiased ability in glycopeptides capture with high sensitivity and selectivity, but also highly effective for phosphopeptides capture. More importantly, the promising nanoprobe enables the simultaneous capture and analysis of glycosylated and phosphorylated peptides from mouse brain, as well as human eye lens of normal and cataract. The identification of glycosylated and phosphorylated sites within normal and cataract reflects their differential expression and offers an excellent insight to glycosylation/phosphorylation-related human eye diseases, indicating its great potential in glycoproteomics and phosphoproteomics researches. A promising nanoprobe functionalized with L-cysteine and titanium ion was fabricated for capture of glycopeptides and phosphopeptides. [Display omitted] •A promising nanoprobe functionalized with l-cysteine and titanium ion was fabricated.•The nanoprobe was used to capture glycopeptides and phosphopeptides respectively and simultaneously.•Glycopeptides and phosphopeptides were identified from mouse brain and human lens of normal and cataract.