Shotgun Glycomics: A Microarray Strategy for Functional Glycomics
| Autor: | David F. Smith, Hong Ju, Baoyun Xia, Chunmei Zhao, Yi Lasanajak, Jamie Heimburg-Molinaro, Xuezheng Song, Ross J. Molinaro, Richard D. Cummings, Jeanne M. Rhea |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Glycan
Erythrocytes Microarray glycan array Chemical biology Shotgun Biology 01 natural sciences Biochemistry Article Cell Line Glycomics 03 medical and health sciences Animals Humans Molecular Biology 030304 developmental biology Lyme Disease 0303 health sciences Molecular Structure glycosphingolipids 010405 organic chemistry fluorescent labeling Cell Biology Microarray Analysis Glycome functional glycomics 0104 chemical sciences immobilization biology.protein lipids (amino acids peptides and proteins) DNA microarray Biotechnology Fluorescent tag |
| Zdroj: | Nature methods |
| ISSN: | 1548-7105 1548-7091 |
| Popis: | Major challenges of glycomics are to characterize a glycome and identify functional glycans as ligands for glycan-binding proteins (GBPs). To address these issues we developed a general strategy termed shotgun glycomics. We focus on glycosphingolipids (GSLs), a class of glycoconjugates that is challenging to study, recognized by toxins, antibodies and GBPs. We derivatized GSLs extracted from cells with a heterobifunctional fluorescent tag suitable for covalent immobilization. We separated fluorescent GSLs by multidimensional chromatography, quantified them and coupled them to glass slides to create GSL shotgun microarrays. Then we interrogated the microarrays with cholera toxin, antibodies and sera from individuals with Lyme disease to identify biologically relevant GSLs that we subsequently characterized by mass spectrometry. Shotgun glycomics incorporating GSLs and potentially glycoprotein-derived glycans is an approach for accessing the complex glycomes of animal cells and is a strategy for focusing structural analyses on functionally important glycans. |
| Databáze: | OpenAIRE |
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