Glycan Microarray Analysis of P-type Lectins Reveals Distinct Phosphomannose Glycan Recognition

• Type of Material: Article
• Authors: Xuezheng Song, Emory University; Yi Lasanajak, Emory University; Linda J. Olson, Medical College of Wisconsin; Marielle Boonen, Washington University; Nancy M. Dahms, Medical College of Wisconsin; Stuart Kornfeld, Washington University; Richard Cummings, Emory University; David Smith, Emory University
• Language: English
• Date: 2009-12-11
• Publisher: American Society for Biochemistry and Molecular Biology
• Publication Version: Final Published Version
• Copyright Statement: © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
• Final Published Version (URL): http://www.jbc.org/content/284/50/35201
• Title of Journal or Parent Work: Journal of Biological Chemistry
• ISSN: 0021-9258
• Volume: 284
• Issue: 50
• Start Page: 35201
• End Page: 35214
• Grant/Funding Information: This work was supported, in whole or in part, by National Institutes of Health Grants R01DK42667 (to N. M. D.), CA08759 (to S. K.), and GM085448 (to D. F. S.).
• Research Categories: Chemistry, Biochemistry

Abstract

The specificity of the cation-independent and -dependent mannose 6-phosphate receptors (CI-MPR and CD-MPR) for high mannose-type N-glycans of defined structure containing zero, one, or two Man-P-GlcNAc phosphodiester or Man-6-P phosphomonoester residues was determined by analysis on a phosphorylated glycan microarray. Amine-activated glycans were covalently printed on N-hydroxysuccinimide-activated glass slides and interrogated with different concentrations of recombinant CD-MPR or soluble CI-MPR. Neither receptor bound to non-phosphorylated glycans. The CD-MPR bound weakly or undetectably to the phosphodiester derivatives, but strongly to the phosphomonoester-containing glycans with the exception of a single Man7GlcNAc2-R isomer that contained a single Man-6-P residue. By contrast, the CI-MPR bound with high affinity to glycans containing either phospho-mono- or -diesters although, like the CD-MPR, it differentially recognized isomers of phosphorylated Man7GlcNAc2-R. This differential recognition of phosphorylated glycans by the CI- and CD-MPRs has implications for understanding the biosynthesis and targeting of lysosomal hydrolases.