Use of glycan microarrays to explore specificity of glycan-binding proteins.
Microarrays of defined glycans represent a high throughput approach to determining the specificity of lectins, or more generally glycan-binding proteins (GBPs). The utility of a glycan microarray is directly related to the number and variety of the glycans available on the printed surface for interrogation by GBPs. The Consortium for Functional Glycomics (CFG), funded by the National Institute of General Medical Sciences (NIGMS), has generated a glycan microarray available to the public as an investigator-driven resource, where hundreds of GBPs have been analyzed. Here we describe the methods generally used by the CFG to prepare glycan arrays and interrogate them with GBPs. We also describe our new approach to normalizing glycan microarray data derived from concentration-dependent analyses of GBP binding, and the application of this approach with the plant lectin Sambucus nigra agglutinin (SNA-I) and human galectin-8. The use of glycan microarrays with this approach readily generates a prediction of the glycan determinants required for high affinity binding by a GBP.
Smith
David F
DF
Department of Biochemistry, The Glycomics Center, Emory University School of Medicine, Atlanta, Georgia, USA.
Song
Xuezheng
X
Cummings
Richard D
RD
D000818:Animals; D001665:Binding Sites; D002240:Carbohydrate Sequence; D054794:Glycomics / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D037102:Lectins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D011134:Polysaccharides / Q000032:analysis* / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D027924:Sambucus nigra / Q000378:metabolism; D013379:Substrate Specificity
D037102:Lectins; D011134:Polysaccharides