Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry, Peking University , Beijing 100871, P. R. China.
Division of Metrology in Chemistry and Analytical Science, National Institute of Metrology , Beijing 100029, P. R. China.
ACS Appl Mater Interfaces. 2017 Jun 14;9(23):19562-19568. doi: 10.1021/acsami.7b02853. Epub 2017 Jun 1.
Cysteine-functionalized metal-organic framework (MOF) was synthesized via a common and facile two-step method of in situ loading of Au nanoparticles on amino-derived MOF followed by l-cysteine (Cys) immobilization. Owing to the large specific surface area and ultrahigh hydrophilicity of this nanocomposite, excellent performance was observed in the enrichment of N-linked glycopeptides in both model glycoprotein and HeLa cell lysate. By using this nanocomposite, 16 and 31 glycopeptides were efficiently extracted from digest of horseradish peroxidase (HRP) and human serum immunoglobulin G (IgG), respectively. The short incubation time (5 min), large binding capacity (150 mg/g, IgG digest to material), good selectivity (1:50, molar ratio of IgG and bovine serum albumin (BSA) digest), high recovery (over 80%), and low detection limit (1 fmol) ensure the effectiveness and robustness of MIL-101(NH)@Au-Cys in complex HeLa cell lysate. As a result, 1123 N-glycosylation sites corresponding to 1069 N-glycopeptides and 614 N-glycoproteins were identified from the lysate. Compared with those of previously reported hydrophilic methods, to our knowledge, it was the best result. This work paves a new way for fast functionalization of MOF and also provides a novel idea for material design in sample preparation, especially in glycoproteome and related analysis.
半胱氨酸功能化金属有机骨架（MOF）通过原位负载金纳米粒子于衍生自氨基酸的 MOF 上，然后再固定 L-半胱氨酸（Cys）的通用且简便的两步法合成。由于该纳米复合材料具有大的比表面积和超高的亲水性，因此在模型糖蛋白和 HeLa 细胞裂解物中的 N-连接糖肽的富集方面表现出优异的性能。使用该纳米复合材料，分别从辣根过氧化物酶（HRP）和人血清免疫球蛋白 G（IgG）的消化物中有效地提取了 16 和 31 种糖肽。短的孵育时间（5 分钟）、大的结合容量（150mg/g，IgG 消化物与材料的摩尔比）、良好的选择性（1:50，IgG 和牛血清白蛋白（BSA）消化物的摩尔比）、高回收率（超过 80%）和低检测限（1fmol）确保了 MIL-101（NH）@Au-Cys 在复杂 HeLa 细胞裂解物中的有效性和稳健性。结果，从裂解物中鉴定出 1123 个对应于 1069 个 N-糖肽和 614 个 N-糖蛋白的 N-糖基化位点。据我们所知，与以前报道的亲水方法相比，这是最好的结果。这项工作为 MOF 的快速功能化开辟了新途径，也为样品制备中的材料设计提供了新的思路，特别是在糖蛋白质组学和相关分析中。
