Duong T Bui, Elena N Kitova, Ling Han, Lara K Mahal, John S Klassen
{"title":"Quantifying Protein-Glycan Interactions Using Native Mass Spectrometry.","authors":"Duong T Bui, Elena N Kitova, Ling Han, Lara K Mahal, John S Klassen","doi":"10.1002/mas.21943","DOIUrl":null,"url":null,"abstract":"<p><p>Interactions between glycan-binding proteins (GBPs) and carbohydrates (glycans) are essential to many biological processes relevant to human health and disease. For most GBPs, however, their glycan interactome-the repertoire of glycans recognized and their specificities-is poorly defined. The structural diversity of biologically relevant glycans and their limited availability in purified form, as well as their varied presentation, often as glycoconjugates, and weak affinities are key challenges hindering comprehensive glycan interaction mapping. Native mass spectrometry (nMS), a versatile, sensitive and label-free tool for the discovery of GBP-glycan interactions and quantifying their stoichiometry and thermodynamic parameters, is poised to play a leading role in defining the glycan interactome of GBPs. Here, we review established nMS methodologies, as well as important experimental and instrumental considerations, for detecting GBP-glycan interactions in vitro, and reliably measuring their stoichiometry and affinity. Recent advances in nMS methods for high-throughput library screening, including shotgun glycomics, and quantifying GBP interactions with glycoproteins and glycosphingolipids, are also described.</p>","PeriodicalId":206,"journal":{"name":"Mass Spectrometry Reviews","volume":" ","pages":""},"PeriodicalIF":6.6000,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mass Spectrometry Reviews","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1002/mas.21943","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"SPECTROSCOPY","Score":null,"Total":0}
引用次数: 0
Abstract
Interactions between glycan-binding proteins (GBPs) and carbohydrates (glycans) are essential to many biological processes relevant to human health and disease. For most GBPs, however, their glycan interactome-the repertoire of glycans recognized and their specificities-is poorly defined. The structural diversity of biologically relevant glycans and their limited availability in purified form, as well as their varied presentation, often as glycoconjugates, and weak affinities are key challenges hindering comprehensive glycan interaction mapping. Native mass spectrometry (nMS), a versatile, sensitive and label-free tool for the discovery of GBP-glycan interactions and quantifying their stoichiometry and thermodynamic parameters, is poised to play a leading role in defining the glycan interactome of GBPs. Here, we review established nMS methodologies, as well as important experimental and instrumental considerations, for detecting GBP-glycan interactions in vitro, and reliably measuring their stoichiometry and affinity. Recent advances in nMS methods for high-throughput library screening, including shotgun glycomics, and quantifying GBP interactions with glycoproteins and glycosphingolipids, are also described.
期刊介绍:
The aim of the journal Mass Spectrometry Reviews is to publish well-written reviews in selected topics in the various sub-fields of mass spectrometry as a means to summarize the research that has been performed in that area, to focus attention of other researchers, to critically review the published material, and to stimulate further research in that area.
The scope of the published reviews include, but are not limited to topics, such as theoretical treatments, instrumental design, ionization methods, analyzers, detectors, application to the qualitative and quantitative analysis of various compounds or elements, basic ion chemistry and structure studies, ion energetic studies, and studies on biomolecules, polymers, etc.