Filter-entrapment enrichment pull-down assay for glycosaminoglycan structural characterization and protein interaction

Authors
Yu, Yanlei
Zhang, Fuming
Renois-Predelus, Gina
Amster, I. Jonathan
Linhardt, Robert J.
ORCID
https://orcid.org/0000-0003-2219-5833
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Issue Date
2020-10-01
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Full Citation
Filter-entrapment enrichment pull-down assay for glycosaminoglycan structural characterization and protein interaction, Y. Yu, F. Zhang, G. Renois-Predelus, I. J. Amster, R. J. Linhardt, Carbohydrate Polymers, 245, 116623, 2020.
Abstract
Heparins are the most pharmaceutically important polysaccharides. These heparin-based anticoagulant/antithrombotic agents include unfractionated heparins, low molecular weight heparins (LMWHs) and ultralow molecular weight heparins (ULMWHs). Heparins exhibit their pharmacological and biological activities through interaction with heparin-binding proteins. The prototypical heparin-binding protein is antithrombin III (AT), responsible for heparin's anticoagulant/antithrombotic activity. This study describes a filter-trapping method to isolate the chains in enoxaparin, a LMWH, which bind to AT. We demonstrate this method using the ULMWH, fondaparinux, which consists of a single well defined AT binding site. The interacting chains of enoxaparin are then characterized by activity assays, top-down liquid chromatography-mass spectrometry, and capillary zone electrophoresis mass spectrometry. This filter-trapping assay is an improvement over affinity chromatography for isolating heparin chains interacting with heparin binding proteins.
Description
Carbohydrate Polymers, 245, 116623
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Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Elsevier
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Carbohydrate Polymers
https://harc.rpi.edu/
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A full text version is available in DSpace@RPI