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Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells

Patnaik, SK; Potvin, B; Nordenfelt, Susanne LU ; Sturm, D; Leffler, Hakon LU and Stanley, P (2006) In Glycobiology 16(4). p.305-317
Abstract
Galectins are implicated in a large variety of biological functions, many of which depend on their carbohydrate-binding ability. Fifteen members of the family have been identified in vertebrates based on binding to galactose (Gal) that is mediated by one or two, evolutionarily conserved, carbohydrate-recognition domains (CRDs). Variations in glycan structures expressed on glycoconjugates at the cell surface may, therefore, affect galectin binding and functions. To identify roles for different glycans in the binding of the three types of mammalian galectins to cells, we performed fluorescence cytometry at 4deg;C with recombinant rat galectin-1, human galectin-3, and three forms of human galectin-8, to Chinese hamster ovary (CHO) cells and... (More)
Galectins are implicated in a large variety of biological functions, many of which depend on their carbohydrate-binding ability. Fifteen members of the family have been identified in vertebrates based on binding to galactose (Gal) that is mediated by one or two, evolutionarily conserved, carbohydrate-recognition domains (CRDs). Variations in glycan structures expressed on glycoconjugates at the cell surface may, therefore, affect galectin binding and functions. To identify roles for different glycans in the binding of the three types of mammalian galectins to cells, we performed fluorescence cytometry at 4deg;C with recombinant rat galectin-1, human galectin-3, and three forms of human galectin-8, to Chinese hamster ovary (CHO) cells and 12 different CHO glycosylation mutants. All galectin species bound to parent CHO cells and binding was inhibited > 90% by 0.2 M lactose. Galectin-8 isoforms with either a long or a short inter-CRD linker bound similarly to CHO cells. However, a truncated form of galectin-8 containing only the N-terminal CRD bound only weakly to CHO cells and the C-terminal galectin-8 CRD exhibited extremely low binding. Binding of the galectins to the different CHO glycosylation mutants revealed that complex N-glycans are the major ligands for each galectin except the N-terminal CRD of galectins-8, and also identified some fine differences in glycan recognition. Interestingly, increased binding of galectin-1 at 4deg;C correlated with increased propidium iodide (PI) uptake, whereas galectin-3 or -8 binding did not induce permeability to PI. The CHO glycosylation mutants with various repertoires of cell surface glycans are a useful tool for investigating galectin-cell interactions as they present complex and simple glycans in a natural mixture of multivalent protein and lipid glycoconjugates anchored in a cell membrane. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
CHO, glycosylation mutants, glycan recognition, lectin, galectin
in
Glycobiology
volume
16
issue
4
pages
305 - 317
publisher
Oxford University Press
external identifiers
  • pmid:16319083
  • wos:000236106200005
  • scopus:33645102970
ISSN
1460-2423
DOI
10.1093/glycob/cwj063
language
English
LU publication?
yes
id
290275d7-8cd1-4654-968a-2cd26adb29ce (old id 415728)
date added to LUP
2007-10-13 18:45:06
date last changed
2019-03-24 04:05:42
@article{290275d7-8cd1-4654-968a-2cd26adb29ce,
  abstract     = {Galectins are implicated in a large variety of biological functions, many of which depend on their carbohydrate-binding ability. Fifteen members of the family have been identified in vertebrates based on binding to galactose (Gal) that is mediated by one or two, evolutionarily conserved, carbohydrate-recognition domains (CRDs). Variations in glycan structures expressed on glycoconjugates at the cell surface may, therefore, affect galectin binding and functions. To identify roles for different glycans in the binding of the three types of mammalian galectins to cells, we performed fluorescence cytometry at 4deg;C with recombinant rat galectin-1, human galectin-3, and three forms of human galectin-8, to Chinese hamster ovary (CHO) cells and 12 different CHO glycosylation mutants. All galectin species bound to parent CHO cells and binding was inhibited > 90% by 0.2 M lactose. Galectin-8 isoforms with either a long or a short inter-CRD linker bound similarly to CHO cells. However, a truncated form of galectin-8 containing only the N-terminal CRD bound only weakly to CHO cells and the C-terminal galectin-8 CRD exhibited extremely low binding. Binding of the galectins to the different CHO glycosylation mutants revealed that complex N-glycans are the major ligands for each galectin except the N-terminal CRD of galectins-8, and also identified some fine differences in glycan recognition. Interestingly, increased binding of galectin-1 at 4deg;C correlated with increased propidium iodide (PI) uptake, whereas galectin-3 or -8 binding did not induce permeability to PI. The CHO glycosylation mutants with various repertoires of cell surface glycans are a useful tool for investigating galectin-cell interactions as they present complex and simple glycans in a natural mixture of multivalent protein and lipid glycoconjugates anchored in a cell membrane.},
  author       = {Patnaik, SK and Potvin, B and Nordenfelt, Susanne and Sturm, D and Leffler, Hakon and Stanley, P},
  issn         = {1460-2423},
  keyword      = {CHO,glycosylation mutants,glycan recognition,lectin,galectin},
  language     = {eng},
  number       = {4},
  pages        = {305--317},
  publisher    = {Oxford University Press},
  series       = {Glycobiology},
  title        = {Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells},
  url          = {http://dx.doi.org/10.1093/glycob/cwj063},
  volume       = {16},
  year         = {2006},
}