Carbohydrate arrays for the evaluation of protein binding and enzymatic modification.
This paper reports a chemical strategy for preparing carbohydrate arrays and utilizes these arrays for the characterization of carbohydrate-protein interactions. Carbohydrate chips were prepared by the Diels-Alder-mediated immobilization of carbohydrate-cyclopentadiene conjugates to self-assembled monolayers that present benzoquinone and penta(ethylene glycol) groups. Surface plasmon resonance spectroscopy showed that lectins bound specifically to immobilized carbohydrates and that the glycol groups prevented nonspecific protein adsorption. Carbohydrate arrays presenting ten monosaccharides were then evaluated by profiling the binding specificities of several lectins. These arrays were also used to determine the inhibitory concentrations of soluble carbohydrates for lectins and to characterize the substrate specificity of beta-1,4-galactosyltransferase. Finally, a strategy for preparing arrays with carbohydrates generated on solid phase is shown. This surface engineering strategy will permit the preparation and evaluation of carbohydrate arrays that present diverse and complex structures.
Houseman
Benjamin T
BT
Department of Chemistry and The Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637, USA.
Mrksich
Milan
M
D001667:Binding, Competitive; D050260:Carbohydrate Metabolism; D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; D003517:Cyclopentanes; D004798:Enzymes / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism; D008832:Microchemistry / Q000379:methods*; D009237:N-Acetyllactosamine Synthase / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; D003517:Cyclopentanes; D004798:Enzymes; D037102:Lectins; D011506:Proteins; D009237:N-Acetyllactosamine Synthase