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

Santosh Kumar Patnaik, Barry Potvin, Susanne Carlsson, David Sturm, Hakon Leffler, Pamela Stanley

Research output: Contribution to journalArticle

100 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)305-317
Number of pages13
JournalGlycobiology
Volume16
Issue number4
DOIs
StatePublished - Apr 2006

Fingerprint

Galectin 1
Galectins
Galectin 3
Cricetulus
Polysaccharides
Ovary
Cells
Ligands
Carbohydrates
Glycosylation
Glycoconjugates
Propidium
Lipid-Linked Proteins
Aptitude
Cell membranes
Lactose
Galactose
Cell Communication
Vertebrates
Rats

Keywords

  • CHO
  • Galectin
  • Glycan recognition
  • Glycosylation mutants
  • Lectin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells. / Patnaik, Santosh Kumar; Potvin, Barry; Carlsson, Susanne; Sturm, David; Leffler, Hakon; Stanley, Pamela.

In: Glycobiology, Vol. 16, No. 4, 04.2006, p. 305-317.

Research output: Contribution to journalArticle

Patnaik, Santosh Kumar ; Potvin, Barry ; Carlsson, Susanne ; Sturm, David ; Leffler, Hakon ; Stanley, Pamela. / Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells. In: Glycobiology. 2006 ; Vol. 16, No. 4. pp. 305-317.
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