Profiling the sulfation specificities of glycosaminoglycan interactions with growth factors and chemotactic proteins using microarrays.
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.
Shipp
Eric L
EL
Division of Chemistry and Chemical Engineering and Howard Hughes Medical Institute, California Institute of Technology, Pasadena, CA 91125, USA.
Hsieh-Wilson
Linda C
LC
D000818:Animals; D001369:Axons / Q000378:metabolism / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002630:Chemotactic Factors / Q000378:metabolism*; D002645:Chickens; D002809:Chondroitin Sulfates / Q000378:metabolism*; D036382:Ephrin-A1 / Q000378:metabolism; D036386:Ephrin-A5 / Q000378:metabolism; D005260:Female; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D046228:Microarray Analysis / Q000379:methods; D009414:Nerve Growth Factors / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D000075388:Netrin-1; D011247:Pregnancy; D011485:Protein Binding; D051381:Rats; D039961:Semaphorins / Q000378:metabolism; D013379:Substrate Specificity; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D000098552:Slit Homolog 2 Protein
D002630:Chemotactic Factors; D002809:Chondroitin Sulfates; D036382:Ephrin-A1; D036386:Ephrin-A5; D005346:Fibroblast Growth Factors; D006497:Heparan Sulfate; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D000075388:Netrin-1; D039961:Semaphorins; D025521:Tumor Suppressor Proteins; D000098552:Slit Homolog 2 Protein; C000617351:NTN1 protein, human; C409766:SEMA3B protein, human