Abstract
We report a carbohydrate microarray-based approach for the rapid, facile analysis of glycosaminoglycan-protein interactions. The key structural determinants responsible for protein binding, such as sulfate groups that participate in the interactions, were elucidated. Specificities were also readily compared across protein families or functional classes, and comparisons among glycosaminoglycan subclasses provided a more comprehensive understanding of protein specificity. To validate the approach, we showed that fibroblast growth factor family members have distinct sulfation preferences. We also demonstrated that heparan sulfate and chondroitin sulfate interact in a sulfation-dependent manner with various axon guidance proteins, including slit2, netrin1, ephrinA1, ephrinA5, and semaphorin5B. We anticipate that these microarrays will accelerate the discovery of glycosaminoglycan-binding proteins and provide a deeper understanding of their roles in regulating diverse biological processes.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Animals
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Axons / metabolism
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Axons / physiology
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Cell Movement / physiology
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Chemotactic Factors / metabolism*
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Chickens
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Chondroitin Sulfates / metabolism*
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Ephrin-A1 / metabolism
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Ephrin-A5 / metabolism
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Female
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Fibroblast Growth Factors / metabolism*
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Heparitin Sulfate / metabolism*
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Intercellular Signaling Peptides and Proteins / metabolism
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Membrane Glycoproteins / metabolism
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Microarray Analysis / methods
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Nerve Growth Factors / metabolism
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Nerve Tissue Proteins / metabolism
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Netrin-1
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Pregnancy
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Protein Binding
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Rats
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Semaphorins / metabolism
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Substrate Specificity
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Tumor Suppressor Proteins / metabolism
Substances
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Chemotactic Factors
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Ephrin-A1
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Ephrin-A5
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Intercellular Signaling Peptides and Proteins
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Membrane Glycoproteins
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NTN1 protein, human
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Nerve Growth Factors
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Nerve Tissue Proteins
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SEMA3B protein, human
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Semaphorins
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Tumor Suppressor Proteins
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Netrin-1
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Fibroblast Growth Factors
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Chondroitin Sulfates
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Heparitin Sulfate
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Slit homolog 2 protein