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Affinity of galectin-8 and its carbohydrate recognition domains for ligands in solution and at the cell surface.

Nordenfelt, Susanne LU ; Öberg, Christopher LU ; Carlsson, Michael C; Sundin, Anders LU ; Nilsson, Ulf LU ; Smith, David; Cummings, Richard D; Almkvist, Jenny; Karlsson, Anna and Leffler, Hakon LU (2007) In Glycobiology 17(6). p.663-676
Abstract
Galectin-8 has two different carbohydrate recognition domains (CRDs), the N-terminal Gal-8N and the C-terminal Gal-8C linked by a peptide, and has various effects on cell adhesion and signaling. To understand the mechanism for these effects further, we compared the binding activities of galectin-8 in solution with its binding and activation of cells. We used glycan array analysis to broaden the specificity profile of the two galectin-8 CRDs, as well as intact galectin-8s (short and long linker), confirming the unique preference for sulfated and sialylated glycans of Gal-8N. Using a fluorescence anisotropy assay, we examined the solution affinities for a subset of these glycans, the highest being 50 nM for NeuAc alpha 2,3Lac by Gal-8N.... (More)
Galectin-8 has two different carbohydrate recognition domains (CRDs), the N-terminal Gal-8N and the C-terminal Gal-8C linked by a peptide, and has various effects on cell adhesion and signaling. To understand the mechanism for these effects further, we compared the binding activities of galectin-8 in solution with its binding and activation of cells. We used glycan array analysis to broaden the specificity profile of the two galectin-8 CRDs, as well as intact galectin-8s (short and long linker), confirming the unique preference for sulfated and sialylated glycans of Gal-8N. Using a fluorescence anisotropy assay, we examined the solution affinities for a subset of these glycans, the highest being 50 nM for NeuAc alpha 2,3Lac by Gal-8N. Thus, carbohydrate-protein interactions can be of high affinity without requiring multivalency. More importantly, using fluorescence polarization, we also gained information on how the affinity is built by multiple weak interactions between different fragments of the glycan and its carrier molecule and the galectin CRD subsites (A-E). In intact galectin-8 proteins, the two domains act independently of each other in solution, whereas at a surface they act together. Ligands with moderate or weak affinity for the isolated CRI)s on the array are bound strongly by intact galectin-8s. Also galectin-8 binding and signaling at cell surfaces can be explained by combined binding of the two CRI)s to low or medium affinity ligands, and their highest affinity ligands, such as sialylated galactosides, are not required. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cell surface, affinity, sialic acid, galectin, specificity
in
Glycobiology
volume
17
issue
6
pages
663 - 676
publisher
Oxford University Press
external identifiers
  • wos:000247678700013
  • scopus:34447322293
ISSN
1460-2423
DOI
10.1093/glycob/cwm026
language
English
LU publication?
yes
id
9018fc84-baed-4fb6-8c8f-2a68005176b7 (old id 166797)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17339281&dopt=Abstract
date added to LUP
2007-07-04 08:33:38
date last changed
2017-10-01 04:53:33
@article{9018fc84-baed-4fb6-8c8f-2a68005176b7,
  abstract     = {Galectin-8 has two different carbohydrate recognition domains (CRDs), the N-terminal Gal-8N and the C-terminal Gal-8C linked by a peptide, and has various effects on cell adhesion and signaling. To understand the mechanism for these effects further, we compared the binding activities of galectin-8 in solution with its binding and activation of cells. We used glycan array analysis to broaden the specificity profile of the two galectin-8 CRDs, as well as intact galectin-8s (short and long linker), confirming the unique preference for sulfated and sialylated glycans of Gal-8N. Using a fluorescence anisotropy assay, we examined the solution affinities for a subset of these glycans, the highest being 50 nM for NeuAc alpha 2,3Lac by Gal-8N. Thus, carbohydrate-protein interactions can be of high affinity without requiring multivalency. More importantly, using fluorescence polarization, we also gained information on how the affinity is built by multiple weak interactions between different fragments of the glycan and its carrier molecule and the galectin CRD subsites (A-E). In intact galectin-8 proteins, the two domains act independently of each other in solution, whereas at a surface they act together. Ligands with moderate or weak affinity for the isolated CRI)s on the array are bound strongly by intact galectin-8s. Also galectin-8 binding and signaling at cell surfaces can be explained by combined binding of the two CRI)s to low or medium affinity ligands, and their highest affinity ligands, such as sialylated galactosides, are not required.},
  author       = {Nordenfelt, Susanne and Öberg, Christopher and Carlsson, Michael C and Sundin, Anders and Nilsson, Ulf and Smith, David and Cummings, Richard D and Almkvist, Jenny and Karlsson, Anna and Leffler, Hakon},
  issn         = {1460-2423},
  keyword      = {cell surface,affinity,sialic acid,galectin,specificity},
  language     = {eng},
  number       = {6},
  pages        = {663--676},
  publisher    = {Oxford University Press},
  series       = {Glycobiology},
  title        = {Affinity of galectin-8 and its carbohydrate recognition domains for ligands in solution and at the cell surface.},
  url          = {http://dx.doi.org/10.1093/glycob/cwm026},
  volume       = {17},
  year         = {2007},
}