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Fluorescence polarization as an analytical tool to evaluate galectin-ligand interactions.

Sörme, Pernilla LU ; Kahl-Knutsson, Barbro; Huflejt, Margaret; Nilsson, Ulf LU and Leffler, Hakon LU (2004) In Analytical Biochemistry 334(1). p.36-47
Abstract
Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of... (More)
Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of nonfluorescent compounds. As the assay is in solution and does not require separation of free and bound probe, it is simple and rapid and can easily be applied to different unlabeled galectins. As all interaction components are known, Kd values for galectin–inhibitor interaction can be directly calculated without approximation other than the assumption of a simple one-site competition. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Biochemistry
volume
334
issue
1
pages
36 - 47
publisher
Elsevier
external identifiers
  • wos:000224710600004
  • pmid:15464951
  • scopus:4644260095
ISSN
1096-0309
DOI
10.1016/j.ab.2004.06.042
language
English
LU publication?
yes
id
b61bce62-d583-449d-bab6-2fa27d301ed0 (old id 129925)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15464951&dopt=Abstract
date added to LUP
2007-07-04 11:39:18
date last changed
2017-07-30 03:30:50
@article{b61bce62-d583-449d-bab6-2fa27d301ed0,
  abstract     = {Galectins are a family of β-galactose binding lectins associated with functions such as immunological and malignant events. To study the binding affinity of galectins for natural and artificial saccharides and glycoconjugates we have developed an assay using fluorescence polarization. A collection of fluorescein-conjugated saccharides was synthesized and used as probes with galectins-1 and -3 and the two carbohydrate recognition domains of galectin-4. Direct binding of a fixed probe amount with different amounts of each galectin defined specificity and selectivity and permitted selection of the optimal probe for inhibition studies. Then fixed amounts of galectin and selected probe were used to screen the inhibitory potency of a library of nonfluorescent compounds. As the assay is in solution and does not require separation of free and bound probe, it is simple and rapid and can easily be applied to different unlabeled galectins. As all interaction components are known, Kd values for galectin–inhibitor interaction can be directly calculated without approximation other than the assumption of a simple one-site competition.},
  author       = {Sörme, Pernilla and Kahl-Knutsson, Barbro and Huflejt, Margaret and Nilsson, Ulf and Leffler, Hakon},
  issn         = {1096-0309},
  language     = {eng},
  number       = {1},
  pages        = {36--47},
  publisher    = {Elsevier},
  series       = {Analytical Biochemistry},
  title        = {Fluorescence polarization as an analytical tool to evaluate galectin-ligand interactions.},
  url          = {http://dx.doi.org/10.1016/j.ab.2004.06.042},
  volume       = {334},
  year         = {2004},
}