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Nat Methods. 2018 Nov;15(11):889-899. doi: 10.1038/s41592-018-0189-6. Epub 2018 Oct 30.

A mutant-cell library for systematic analysis of heparan sulfate structure-function relationships.

Author information

1
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.
2
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.
3
Institute of Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
4
Departments of Chemistry and Chemical Biology, Chemical and Biological Engineering, and Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
5
Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Ocean University of China, Qingdao, China.
6
Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Nijmegen, the Netherlands.
7
Division of Pulmonary and Critical Care Medicine, Department of Medicine, Columbia University Medical Center, New York, NY, USA.
8
Multidisciplinary Pain Center, Aichi Medical University, Nagakute, Japan.
9
Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
10
Glycobiology Research and Training Center, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
11
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA. Lwang@ccrc.uga.edu.
12
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA. Lwang@ccrc.uga.edu.

Abstract

Heparan sulfate (HS) is a complex linear polysaccharide that modulates a wide range of biological functions. Elucidating the structure-function relationship of HS has been challenging. Here we report the generation of an HS-mutant mouse lung endothelial cell library by systematic deletion of HS genes expressed in the cell. We used this library to (1) determine that the strictly defined fine structure of HS, not its overall degree of sulfation, is more important for FGF2-FGFR1 signaling; (2) define the epitope features of commonly used anti-HS phage display antibodies; and (3) delineate the fine inter-regulation networks by which HS genes modify HS and chain length in mammalian cells at a cell-type-specific level. Our mutant-cell library will allow robust and systematic interrogation of the roles and related structures of HS in a cellular context.

PMID:
30377379
PMCID:
PMC6214364
[Available on 2019-04-30]
DOI:
10.1038/s41592-018-0189-6

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