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N-Glycans on the link domain of human HARE/Stabilin-2 are needed for hyaluronan binding to purified ecto-domain, but not for cellular endocytosis of hyaluronan.
- Source :
-
Glycobiology . Aug2010, Vol. 20 Issue 8, p991-1001. 11p. 1 Diagram, 5 Graphs. - Publication Year :
- 2010
-
Abstract
- The hyaluronic acid receptor for endocytosis (HARE)/Stabilin-2 is the primary systemic scavenger receptor for 13 ligands including hyaluronan (HA), heparin and chondroitin sulfates. Most ligand-binding sites are within the 190 kDa isoform, which contains ∼25 kDa of N-glycans and is the C-terminal half of the full-length 315 kDa HARE. Glycoproteomic analyses of purified recombinant human 190-HARE ecto-domain identified a diverse population of glycans at 10 of 17 consensus sites. The most diversity (and the only sialylated structures) occurred at N2280, within the HA-binding Link domain. To determine if these N-glycans are required for HA binding, we created human Flp-In 293 cell lines expressing membrane-bound or soluble ecto-domain variants of 190-HARE(N2280A). Membrane-bound HARE lacking Link domain N-glycans mediated rapid HA endocytosis, but purified 190-HARE(N2280A) ecto-domain showed little or no HA binding in ELISA-like, HA-HARE pull-down assays or by surface plasmon resonance analysis (which detected very high apparent affinity for 190-HARE ecto-domain binding to HA; Kd = 5.2 nM). The results indicate that Link domain N-glycans stabilize interactions that facilitate HA binding to HARE. [ABSTRACT FROM PUBLISHER]
- Subjects :
- *HYALURONIC acid
*ENDOCYTOSIS
*HEPARIN
*CHONDROITIN
*PROTEIN binding
Subjects
Details
- Language :
- English
- ISSN :
- 09596658
- Volume :
- 20
- Issue :
- 8
- Database :
- Academic Search Index
- Journal :
- Glycobiology
- Publication Type :
- Academic Journal
- Accession number :
- 51958371
- Full Text :
- https://doi.org/10.1093/glycob/cwq057