基于双功能荧光标签8-氨基1,3,6-萘三磺酸的N-连接糖基化富集方法

  • 乔晓强 ,
  • 王蕊 ,
  • 马宝伟
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  • 河北大学药学院,河北省药物质量研究重点实验室,河北保定071002

网络出版日期: 2013-09-25

基金资助

国家自然科学基金资助项目,河北省自然科学基金资助项目,河北省科学技术研究与发展计划项目,河北大学自然科学研究计划项目

Exploring the N-linked glycosylation capture method based on bio-functional fluorescent tag 8-aminonaphthalene-1,3,6-trisulfonic acid disodium salt

  • QIAO Xiaoqiang ,
  • WANG Rui ,
  • MA Baowei
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Online published: 2013-09-25

摘要

发展了基于双功能荧光标签8-氨基1,3,6-萘三磺酸(ANTS)选择性糖基化蛋白质标记和二氧化钛(TiO2)亲和富集的N-连接糖基化富集新方法.模式蛋白质细胞色素c、肌红蛋白、核糖核酸酶b、抗生素和α-l-抗胰蛋白酶经ANTS标记后,所有糖基化蛋白质均发出亮绿色荧光,而非糖基化蛋白质则不能观测到荧光;进一步采用TiO2对酶切产生的N-连接糖基化肽段进行富集,同时实现了中性糖肽和唾液酸糖肤的选择性富集.通过进一步分析人血清N-连接糖基化蛋白质,共鉴定到39个唯一的N-连接糖蛋白,包括了53个唯一的N-连接糖基化位点;此外,该方法与采用TiO2法直接富集以及肼化学法富集鉴定到的交叠糖基化位点数均小于40%,说明这一新方法与传统方法在糖基化肽段富集方面具有一定的互补性,有望用于复杂蛋白质样品的N-连接糖基化高效富集.

本文引用格式

乔晓强 , 王蕊 , 马宝伟 . 基于双功能荧光标签8-氨基1,3,6-萘三磺酸的N-连接糖基化富集方法[J]. 河北大学学报(自然科学版), 2013 , 33(5) : 489 -495 . DOI: 10.3969/j.issn.1000-1565.2013.05.009

参考文献

[1] 梁鑫淼, LIANG Xinmiao. 基于亲水作用色谱的寡糖色谱分离新进展 [J]. 色谱 2011.doi:10.3724/SP.J.1123.2011.00191
[2] 卢庄, 王京兰, 贾伟, 杨兵, 蔡耘, 邓玉林, 张玉奎, 钱小红, LU Zhuang, WANG Jinglan, JIA Wei, YANG Bing, CAI Yun, DENG Yulin, ZHANG Yukui, QIAN Xiaohong. 麦胚凝集素亲和色谱富集糖肽过程中的肽键裂解 [J]. 色谱 2009.doi:10.3321/j.issn:1000-8713.2009.01.003
[3] 马海宁, 华玉娟, 屠春燕, 袁丽红, 韦萍, MA Haining, HUA Yujuan, TU Chunyan, YUAN Lihong, WEI Ping. 毛细管电泳法分析藏红花植物细胞多糖中单糖组成 [J]. 色谱 2012.doi:10.3724/SP.J.1123.2011.11015
[4] Parodi AJ.. Protein glucosylation and its role in protein folding [Review] [J]. Annual Review of Biochemistry 2000, 0(0).
[5] Danielle H. Dube, Carolyn R. Bertozzi. GLYCANS IN CANCER AND INFLAMMATION - POTENTIAL FOR THERAPEUTICS AND DIAGNOSTICS [J]. Nature reviews Drug discovery 2005, 6(6).
[6] SHEPARD H M, LEWIS G D, SARUP J C. Monoclonal antibody therapy of human cancer:taking the HER2 protooncogene to the clinic [J]. {H}Journal of Clinical Immunology 1991, 11(03).
[7] Liu X, Ma L, Li JJ. Recent developments in the enrichment of glycopeptides for glycoproteomics [J]. Analytical Letters 2008, 2(2).
[8] Jun Zhu, Fangjun Wang, Rui Chen, Kai Cheng, Bo Xu, Zhimou Guo, Xinmiao Liang, Mingliang Ye, Hanfa Zou. Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis [J]. Analytical chemistry 2012, 11(11).
[9] Fanayan, S., Hincapie, M., Hancock, W.S.. Using lectins to harvest the plasma/serum glycoproteome [J]. Electrophoresis: The Official Journal of the International Electrophoresis Society 2012, 12(12).
[10] LI Hengye, WANG Heye, LIU Yunchun. A benzoboroxole-functionalized monolithic column for the selective enrichment and separation of cis-diol containing biomolecules [J]. Chemical Communications 2012, 48(34).
[11] Qu, Y., Liu, J., Yang, K., Liang, Z., Zhang, L., Zhang, Y.. Boronic acid functionalized core-shell polymer nanoparticles prepared by distillation precipitation polymerization for glycopeptide enrichment [J]. Chemistry: A European journal 2012, 29(29).
[12] KIM Y J, ZAIDI AINOUCH Z, GALLIEN S. Mass spec' rometry-based detection and quantification of plasma glycoproteins using selective reaction monitoring [J]. {H}Nature Protocols 2012, 7(05).
[13] Sun B, Ranish JA, Utleg AG, White JT, Yan X, Lin B, Hood L. Shotgun glycopeptide capture approach coupled with mass spectrometry for comprehensive glycoproteomics. [J]. Molecular & cellular proteomics: MCP 2007, 1(1).
[14] PAN Sheng, CHEN Ru, AEBERSOLD R. Mass spectrometry based glycoproteomics-from a proteomics perspective [J]. {H}Molecular & Cellular Proteomics 2010.
[15] Cao, J, Shen, CP, Wang, H, Shen, HL, Chen, YC, Nie, AY, Yan, GQ, Lu, HJ, Liu, YK, Yang, PY. Identification of N-Glycosylation Sites on Secreted Proteins of Human Hepatocellular Carcinoma Cells with a Complementary Proteomics Approach [J]. Journal of Proteome Research 2009, 2(2).
[16] Lee, A, Kolarich, D, Haynes, PA, Jensen, PH, Bakert, MS, Packer, NH. Rat Liver Membrane Glycoproteome: Enrichment by Phase Partitioning and Glycoprotein Capture [J]. Journal of Proteome Research 2009, 2(2).
[17] Pan S, Wang Y, Quinn JF, Peskind ER, Waichunas D, Wimberger JT, Jin JH, Li JG, Zhu D, Pan C. Identification of glycoproteins in human cerebrospinal fluid with a complementary proteomic approach [J]. Journal of proteome research 2006, 10(10).
[18] Qiao, X., Sun, L., Wang, L., Liang, Y., Zhang, L., Shan, Y., Peng, X., Liang, Z., Zhang, Y.. High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization [J]. Journal of chromatography, B. Analytical technologies in the biomedical and life sciences 2011, 17/18(17/18).
[19] Larsen MR, Jensen SS, Jakobsen LA, Heegaard NHH. Exploring the sialiome using titanium dioxide chromatography and mass spectrometry [J]. Molecular & cellular proteomics: MCP 2007, 10(10).
[20] Zhang H, Li XJ, Martin DB, Aebersold R. Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry [J]. Nature biotechnology 2003, 6(6).
[21] Qiao X, Tao D, Qu Y, Sun L, Gao L, Zhang X, Liang Z, Zhang L, Zhang Y. Large-scale N-glycoproteome map of rat brain tissue: simultaneous characterization of insoluble and soluble protein fractions. [J]. Proteomics 2011, 21(21).
[22] Chen, R, Jiang, XN, Sun, DG, Han, GH, Wang, FJ, Ye, ML, Wang, LM, Zou, HF. Glycoproteomics Analysis of Human Liver Tissue by Combination of Multiple Enzyme Digestion and Hydrazide Chemistry [J]. Journal of proteome research 2009, 2(2).
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