Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7: Evidence for differential binding specificities

Nisar Ahmad, Hans J. Gabius, Herbert Kaltner, Sabine André, Ichiro Kuwabara, Fu Tong Liu, Stefan Oscarson, Thomas Norberg, Curtis F. Brewer

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

Binding of a series of sialylated and non-sialylated cell surface carbohydrates to bovine heart galectin-1, recombinant murine galectin-3, and recombinant human galectin-7 was investigated by isothermal titration microcalorimetry (ITC) and hemagglutination inhibition measurements. Galectin-7 shows nearly equal affinities for lactose and Galβ (1-4)GlcNAc (LacNAc-II). Galectin-7, however, displays six- and 11-fold weaker affinity for LacNAc-II compared with galectins-1 and -3, respectively. The affinity of galectin-7 for LacNAc-II containing oligosaccharides is also weaker than the other two galectins. ITC measurements show that all three galectins bind to di- and trimeric oligomers of LacNAc-II, which are epitopes found in poly-N-acetyllactosamine chains of glycoprotein receptors, with affinity constants similar to that of LacNAc-II. The binding valencies of the di- and trimeric LacNAc-II oligomers were observed to be one from ITC measurements, indicating formation of 1:1 complexes with all three galectins. Thus, galectins-1, -3, and -7 all possess binding sites that primarily accommodate one LacNAc-II moiety per monomer of protein. Sialylated oligosaccharides show different specificities for the three galectins. While 2,3-sialyl LacNAc-II binds to all three galectins, 2,6-sialyl LacNAc-II fails to bind to any of the galectins; 2,6-sialylated diLacNAc binds well to galectin-3 and galectin-7, but only weakly to galectin-1. Similar results are obtained with 2,6-sialyl lacto-N-neo-tetraose, which has a reducing end lactose moiety. Thus, unlike galectin-1, which predominantly recognizes non-reducing terminal LacNAc-II residues in oligosaccharides, galectins-3 and -7 recognize both non-reducing terminal LacNAc-II residues as well as internal LacNAc-II and lactose residues in sialylated and non-sialylated oligosaccharides.

Original languageEnglish (US)
Pages (from-to)1096-1104
Number of pages9
JournalCanadian Journal of Chemistry
Volume80
Issue number8
DOIs
StatePublished - 2002

Fingerprint

Galectin 1
Galectins
Epitopes
Oligosaccharides
Carbohydrates
Titration
Thermodynamics
Oligomers
Galectin 3
Lactose
Glycoproteins
Binding sites
Monomers
Galectin 2
Proteins
Binding Sites

Keywords

  • Binding specificities
  • Carbohydrates
  • Galectins
  • Isothermal titration microcalorimetry
  • Lectins

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7 : Evidence for differential binding specificities. / Ahmad, Nisar; Gabius, Hans J.; Kaltner, Herbert; André, Sabine; Kuwabara, Ichiro; Liu, Fu Tong; Oscarson, Stefan; Norberg, Thomas; Brewer, Curtis F.

In: Canadian Journal of Chemistry, Vol. 80, No. 8, 2002, p. 1096-1104.

Research output: Contribution to journalArticle

Ahmad, Nisar ; Gabius, Hans J. ; Kaltner, Herbert ; André, Sabine ; Kuwabara, Ichiro ; Liu, Fu Tong ; Oscarson, Stefan ; Norberg, Thomas ; Brewer, Curtis F. / Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7 : Evidence for differential binding specificities. In: Canadian Journal of Chemistry. 2002 ; Vol. 80, No. 8. pp. 1096-1104.
@article{1bcbf5b1f67d4210a97c16cde7d6cafc,
title = "Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7: Evidence for differential binding specificities",
abstract = "Binding of a series of sialylated and non-sialylated cell surface carbohydrates to bovine heart galectin-1, recombinant murine galectin-3, and recombinant human galectin-7 was investigated by isothermal titration microcalorimetry (ITC) and hemagglutination inhibition measurements. Galectin-7 shows nearly equal affinities for lactose and Galβ (1-4)GlcNAc (LacNAc-II). Galectin-7, however, displays six- and 11-fold weaker affinity for LacNAc-II compared with galectins-1 and -3, respectively. The affinity of galectin-7 for LacNAc-II containing oligosaccharides is also weaker than the other two galectins. ITC measurements show that all three galectins bind to di- and trimeric oligomers of LacNAc-II, which are epitopes found in poly-N-acetyllactosamine chains of glycoprotein receptors, with affinity constants similar to that of LacNAc-II. The binding valencies of the di- and trimeric LacNAc-II oligomers were observed to be one from ITC measurements, indicating formation of 1:1 complexes with all three galectins. Thus, galectins-1, -3, and -7 all possess binding sites that primarily accommodate one LacNAc-II moiety per monomer of protein. Sialylated oligosaccharides show different specificities for the three galectins. While 2,3-sialyl LacNAc-II binds to all three galectins, 2,6-sialyl LacNAc-II fails to bind to any of the galectins; 2,6-sialylated diLacNAc binds well to galectin-3 and galectin-7, but only weakly to galectin-1. Similar results are obtained with 2,6-sialyl lacto-N-neo-tetraose, which has a reducing end lactose moiety. Thus, unlike galectin-1, which predominantly recognizes non-reducing terminal LacNAc-II residues in oligosaccharides, galectins-3 and -7 recognize both non-reducing terminal LacNAc-II residues as well as internal LacNAc-II and lactose residues in sialylated and non-sialylated oligosaccharides.",
keywords = "Binding specificities, Carbohydrates, Galectins, Isothermal titration microcalorimetry, Lectins",
author = "Nisar Ahmad and Gabius, {Hans J.} and Herbert Kaltner and Sabine Andr{\'e} and Ichiro Kuwabara and Liu, {Fu Tong} and Stefan Oscarson and Thomas Norberg and Brewer, {Curtis F.}",
year = "2002",
doi = "10.1139/v02-162",
language = "English (US)",
volume = "80",
pages = "1096--1104",
journal = "Canadian Journal of Chemistry",
issn = "0008-4042",
publisher = "National Research Council of Canada",
number = "8",

}

TY - JOUR

T1 - Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7

T2 - Evidence for differential binding specificities

AU - Ahmad, Nisar

AU - Gabius, Hans J.

AU - Kaltner, Herbert

AU - André, Sabine

AU - Kuwabara, Ichiro

AU - Liu, Fu Tong

AU - Oscarson, Stefan

AU - Norberg, Thomas

AU - Brewer, Curtis F.

PY - 2002

Y1 - 2002

N2 - Binding of a series of sialylated and non-sialylated cell surface carbohydrates to bovine heart galectin-1, recombinant murine galectin-3, and recombinant human galectin-7 was investigated by isothermal titration microcalorimetry (ITC) and hemagglutination inhibition measurements. Galectin-7 shows nearly equal affinities for lactose and Galβ (1-4)GlcNAc (LacNAc-II). Galectin-7, however, displays six- and 11-fold weaker affinity for LacNAc-II compared with galectins-1 and -3, respectively. The affinity of galectin-7 for LacNAc-II containing oligosaccharides is also weaker than the other two galectins. ITC measurements show that all three galectins bind to di- and trimeric oligomers of LacNAc-II, which are epitopes found in poly-N-acetyllactosamine chains of glycoprotein receptors, with affinity constants similar to that of LacNAc-II. The binding valencies of the di- and trimeric LacNAc-II oligomers were observed to be one from ITC measurements, indicating formation of 1:1 complexes with all three galectins. Thus, galectins-1, -3, and -7 all possess binding sites that primarily accommodate one LacNAc-II moiety per monomer of protein. Sialylated oligosaccharides show different specificities for the three galectins. While 2,3-sialyl LacNAc-II binds to all three galectins, 2,6-sialyl LacNAc-II fails to bind to any of the galectins; 2,6-sialylated diLacNAc binds well to galectin-3 and galectin-7, but only weakly to galectin-1. Similar results are obtained with 2,6-sialyl lacto-N-neo-tetraose, which has a reducing end lactose moiety. Thus, unlike galectin-1, which predominantly recognizes non-reducing terminal LacNAc-II residues in oligosaccharides, galectins-3 and -7 recognize both non-reducing terminal LacNAc-II residues as well as internal LacNAc-II and lactose residues in sialylated and non-sialylated oligosaccharides.

AB - Binding of a series of sialylated and non-sialylated cell surface carbohydrates to bovine heart galectin-1, recombinant murine galectin-3, and recombinant human galectin-7 was investigated by isothermal titration microcalorimetry (ITC) and hemagglutination inhibition measurements. Galectin-7 shows nearly equal affinities for lactose and Galβ (1-4)GlcNAc (LacNAc-II). Galectin-7, however, displays six- and 11-fold weaker affinity for LacNAc-II compared with galectins-1 and -3, respectively. The affinity of galectin-7 for LacNAc-II containing oligosaccharides is also weaker than the other two galectins. ITC measurements show that all three galectins bind to di- and trimeric oligomers of LacNAc-II, which are epitopes found in poly-N-acetyllactosamine chains of glycoprotein receptors, with affinity constants similar to that of LacNAc-II. The binding valencies of the di- and trimeric LacNAc-II oligomers were observed to be one from ITC measurements, indicating formation of 1:1 complexes with all three galectins. Thus, galectins-1, -3, and -7 all possess binding sites that primarily accommodate one LacNAc-II moiety per monomer of protein. Sialylated oligosaccharides show different specificities for the three galectins. While 2,3-sialyl LacNAc-II binds to all three galectins, 2,6-sialyl LacNAc-II fails to bind to any of the galectins; 2,6-sialylated diLacNAc binds well to galectin-3 and galectin-7, but only weakly to galectin-1. Similar results are obtained with 2,6-sialyl lacto-N-neo-tetraose, which has a reducing end lactose moiety. Thus, unlike galectin-1, which predominantly recognizes non-reducing terminal LacNAc-II residues in oligosaccharides, galectins-3 and -7 recognize both non-reducing terminal LacNAc-II residues as well as internal LacNAc-II and lactose residues in sialylated and non-sialylated oligosaccharides.

KW - Binding specificities

KW - Carbohydrates

KW - Galectins

KW - Isothermal titration microcalorimetry

KW - Lectins

UR - http://www.scopus.com/inward/record.url?scp=0036436051&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036436051&partnerID=8YFLogxK

U2 - 10.1139/v02-162

DO - 10.1139/v02-162

M3 - Article

AN - SCOPUS:0036436051

VL - 80

SP - 1096

EP - 1104

JO - Canadian Journal of Chemistry

JF - Canadian Journal of Chemistry

SN - 0008-4042

IS - 8

ER -