Back to Search
Start Over
Construction of heparan sulfate microarray for investigating the binding of specific saccharide sequences to proteins
- Source :
- Glycobiology
- Publication Year :
- 2020
- Publisher :
- Oxford University Press (OUP), 2020.
-
Abstract
- Heparan sulfate (HS) is a heterogeneous, extracellular glycan that interacts with proteins and other molecules affecting many biological processes. The specific binding motifs of HS interactions are of interest, but have not been extensively characterized. Glycan microarrays are valuable tools that can be used to probe the interactions between glycans and their ligands while relying on relatively small amounts of samples. Recently, chemoenzymatic synthesis of HS has been employed to produce specific HS structures that can otherwise be difficult to produce. In this study, a microarray of diverse chemoenzymatically synthesized HS structures was developed and HS interactions were characterized. Fluorescently labeled antithrombin III (AT) and fibroblast growth factor-2 (FGF2) were screened against 95 different HS structures under three different printing concentrations to confirm the utility of this microarray. Specific sulfation patterns were found to be important for binding to these proteins and results are consistent with previous specificity studies. Furthermore, the binding affinities (KD,surf) of AT and FGF2 to multiple HS structures were determined using a microarray technique and is consistent with previous reports. Lastly, the 95-compound HS microarray was used to determine the distinct binding profiles for interleukin 12 and platelet factor 4. This technique is ideal for rapid expansion and will be pivotal to the high-throughput characterization of biologically important structure/function relationships.
- Subjects :
- Glycan
Microarray
Antithrombin III
010402 general chemistry
01 natural sciences
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Sulfation
Carbohydrate Conformation
medicine
Extracellular
Humans
Fibroblast
030304 developmental biology
0303 health sciences
Binding Sites
biology
Chemistry
Heparan sulfate
Microarray Analysis
0104 chemical sciences
medicine.anatomical_structure
Carbohydrate Sequence
Analytical Glycobiology
biology.protein
Fibroblast Growth Factor 2
Heparitin Sulfate
DNA microarray
Function (biology)
Subjects
Details
- ISSN :
- 14602423
- Volume :
- 31
- Database :
- OpenAIRE
- Journal :
- Glycobiology
- Accession number :
- edsair.doi.dedup.....b9986175b3f0240d8db0db7fc40ccf88