Glycan microarray analysis of P-type lectins reveals distinct phosphomannose glycan recognition.
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.
Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Xuezheng
X
Song
Yi
Y
Lasanajak
Linda J
LJ
Olson
Marielle
M
Boonen
Nancy M
NM
Dahms
Stuart
S
Kornfeld
Richard D
RD
Cummings
David F
DF
Smith
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D037102:Lectins / Q000737:chemistry*; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011134:Polysaccharides / Q000032:analysis*; D020033:Protein Isoforms / Q000378:metabolism*; D017527:Receptor, IGF Type 2 / Q000378:metabolism*
D037102:Lectins; D009844:Oligosaccharides; D011134:Polysaccharides; D020033:Protein Isoforms; D017527:Receptor, IGF Type 2