l-cysteine-modified metal-organic frameworks as multifunctional probes for efficient identification of N-linked glycopeptides and phosphopeptides in human crystalline lens
| Autor: | Chunhui Deng, Qianjing Liu, Yonglei Wu |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Phosphopeptides
Surface Properties 02 engineering and technology Mass spectrometry Proteomics 01 natural sciences Biochemistry Horseradish peroxidase Analytical Chemistry Lens protein Affinity chromatography Lens Crystalline Humans Environmental Chemistry Cysteine Particle Size Metal-Organic Frameworks Spectroscopy biology Chemistry Hydrophilic interaction chromatography 010401 analytical chemistry Glycopeptides 021001 nanoscience & nanotechnology Combinatorial chemistry 0104 chemical sciences Molecular Probes biology.protein Metal-organic framework 0210 nano-technology Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Analytica Chimica Acta. 1061:110-121 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2019.01.052 |
| Popis: | Highly selective enrichment of N-linked glycopeptides and phosphopeptides from complex biological samples is extremely important prior to mass spectrometry analysis due to their low abundance as well as numerous extrinsic interferences. In this work, l -cysteine (L-Cys)-modified multifunctional metal-organic frameworks denoted as Fe3O4@PDA@MIL-125@Au@L-Cys (mMIL-125@Au@L-Cys) were prepared by modifications step by step. By combining hydrophilic interaction chromatography (HILIC) with metal oxide affinity chromatography (MOAC), the as-prepared material was firstly utilized to identify N-linked glycopeptides and phosphopeptides from tryptic digests of horseradish peroxidase (HRP) and beta-casein (β-casein), respectively, with the help of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and exhibited outstanding sensitivity (0.1 fmol μL−1), great reusability (5 circles) and high selectivity (1: 100). Based on this, it was further applied into the enrichment of glycopeptides and phosphopeptides from tryptic digests of 100 μg human crystalline lens proteins. In the end, 81 N-linked glycopeptides corresponding to 35 glycoproteins and 175 phosphopeptides ascribed to 55 phosphorylated proteins were identified, respectively. The remarkable results were benefitted from the merits of improved hydrophilicity from L-Cys, strong affinity of Ti O centers, numerous reaction sites on the large surface of MOFs and superparamagnetism from Fe3O4 cores. The design of mMIL-125@Au@L-Cys not only served as a multifunctional probe for efficient identification of N-linked glycopeptides and phosphopeptides in human crystalline lens, but also set a precedent for fabricating more MOFs with post-modifications for further proteomics research. |
| Databáze: | OpenAIRE |
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