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Interaction of the verotoxin 1B subunit with soluble aminodeoxy analogues of globotriaosyl ceramides

Mylvaganam, M; Hansen, H C; Binnington, B; Magnusson, Göran LU ; Nyholm, PG and Lingwood, CA (2002) In Biochemical Journal 368. p.769-776
Abstract
Specific hydroxy groups of the terminal disaccharide unit of globotriaosyl ceramide (Gb(3)Cer) were identified from binding studies with deoxyGb(3)Cer and verotoxins (VTs) [Nyholm, Magnusson, Zheng, Norel, Birmington-Boyd and Lingwood (1996) Chem. Biol. 3, 263-275]. Four such hydroxy groups (2", 4", 6" and 6') were each substituted with an amino group and the corresponding deoxyamino globotrioses were conjugated to a ceramide-like aglycone which contained an adamantyl group instead of an acyl chain. Such aglycone modification significantly enhanced the water-solubility of the glycoconjugates [Mylvaganam and Lingwood (1999) Biochem. Biophys. Res. Commun. 257, 391-394]. The inhibitory potential of these soluble aminodeoxy conjugates on the... (More)
Specific hydroxy groups of the terminal disaccharide unit of globotriaosyl ceramide (Gb(3)Cer) were identified from binding studies with deoxyGb(3)Cer and verotoxins (VTs) [Nyholm, Magnusson, Zheng, Norel, Birmington-Boyd and Lingwood (1996) Chem. Biol. 3, 263-275]. Four such hydroxy groups (2", 4", 6" and 6') were each substituted with an amino group and the corresponding deoxyamino globotrioses were conjugated to a ceramide-like aglycone which contained an adamantyl group instead of an acyl chain. Such aglycone modification significantly enhanced the water-solubility of the glycoconjugates [Mylvaganam and Lingwood (1999) Biochem. Biophys. Res. Commun. 257, 391-394]. The inhibitory potential of these soluble aminodeoxy conjugates on the binding of VT1 to Gb(3)Cer immobilized on an ELISA plate was evaluated. Only the 2" and the 6' deoxyamino conjugates were effective inhibitors (IC50 10 muM); the 4" and 6" conjugates were ineffective up to 10 mM. To evaluate the importance of incorporating a rigid adamantyl hydrocarbon group into the ceramide aglycone, globotriaose was conjugated to a t-butylacetamido or an adamantaneacetamido aglycone. By similar ELISAs, only the adamantaneacetamido conjugate inhibited the binding of VT1 to Gb(3)Cer. When deoxyamino conjugates were adsorbed to silica on TLC plates, only the 2" and 6" conjugates bound VT1 and VT2. By a similar TLC assay, acetamido derivatives of 2" and 6' deoxyamino conjugates showed less binding to VT1 and VT2. Neither the crystallographically determined structure of the VT1- globotriaose complex nor modelling studies fully explain the binding patterns shown by these deoxyamino glycoconjugates. Enhanced solvation of the ammonium group of the deoxyamino conjugate could enforce greater constraints in the binding interactions. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
mimic, soluble glycolipid, glycolipid receptor, haemolytic uraemic syndrome
in
Biochemical Journal
volume
368
pages
769 - 776
publisher
Portland Press Limited
external identifiers
  • pmid:12175338
  • wos:000180061800011
  • scopus:0346667169
ISSN
0264-6021
DOI
10.1042/BJ20020225
language
English
LU publication?
yes
id
b3243da3-8e14-4b5d-8e80-81391703a15f (old id 321123)
date added to LUP
2007-11-16 09:56:39
date last changed
2017-01-01 07:14:52
@article{b3243da3-8e14-4b5d-8e80-81391703a15f,
  abstract     = {Specific hydroxy groups of the terminal disaccharide unit of globotriaosyl ceramide (Gb(3)Cer) were identified from binding studies with deoxyGb(3)Cer and verotoxins (VTs) [Nyholm, Magnusson, Zheng, Norel, Birmington-Boyd and Lingwood (1996) Chem. Biol. 3, 263-275]. Four such hydroxy groups (2", 4", 6" and 6') were each substituted with an amino group and the corresponding deoxyamino globotrioses were conjugated to a ceramide-like aglycone which contained an adamantyl group instead of an acyl chain. Such aglycone modification significantly enhanced the water-solubility of the glycoconjugates [Mylvaganam and Lingwood (1999) Biochem. Biophys. Res. Commun. 257, 391-394]. The inhibitory potential of these soluble aminodeoxy conjugates on the binding of VT1 to Gb(3)Cer immobilized on an ELISA plate was evaluated. Only the 2" and the 6' deoxyamino conjugates were effective inhibitors (IC50 10 muM); the 4" and 6" conjugates were ineffective up to 10 mM. To evaluate the importance of incorporating a rigid adamantyl hydrocarbon group into the ceramide aglycone, globotriaose was conjugated to a t-butylacetamido or an adamantaneacetamido aglycone. By similar ELISAs, only the adamantaneacetamido conjugate inhibited the binding of VT1 to Gb(3)Cer. When deoxyamino conjugates were adsorbed to silica on TLC plates, only the 2" and 6" conjugates bound VT1 and VT2. By a similar TLC assay, acetamido derivatives of 2" and 6' deoxyamino conjugates showed less binding to VT1 and VT2. Neither the crystallographically determined structure of the VT1- globotriaose complex nor modelling studies fully explain the binding patterns shown by these deoxyamino glycoconjugates. Enhanced solvation of the ammonium group of the deoxyamino conjugate could enforce greater constraints in the binding interactions.},
  author       = {Mylvaganam, M and Hansen, H C and Binnington, B and Magnusson, Göran and Nyholm, PG and Lingwood, CA},
  issn         = {0264-6021},
  keyword      = {mimic,soluble glycolipid,glycolipid receptor,haemolytic uraemic syndrome},
  language     = {eng},
  pages        = {769--776},
  publisher    = {Portland Press Limited},
  series       = {Biochemical Journal},
  title        = {Interaction of the verotoxin 1B subunit with soluble aminodeoxy analogues of globotriaosyl ceramides},
  url          = {http://dx.doi.org/10.1042/BJ20020225},
  volume       = {368},
  year         = {2002},
}