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Mutational tuning of galectin-3 specificity and biological function.

Salomonsson, Emma LU ; Carlsson, Michael LU ; Osla, Veronica; Hendus-Altenburger, Ruth; Kahl Knutson, Barbro LU ; Öberg, Christopher LU ; Sundin, Anders LU ; Nilsson, Rickard; Nordberg Karlsson, Eva LU and Nilsson, Ulf LU , et al. (2010) In Journal of Biological Chemistry 285(45). p.35079-35091
Abstract
Galectins are defined by a conserved beta-galactoside binding site, which has been linked to many of their important functions in e.g. cell adhesion, signaling and intracellular trafficking. Weak adjacent sites may enhance or decrease affinity for natural beta-galactoside containing glycoconjugates, but little is known about the biological role of this modulation of affinity (fine specificity). We have now produced 10 mutants of human galectin-3, with changes in these adjacent sites, which have altered carbohydrate-binding fine specificity but which retain the basic beta-galactoside binding activity as show by glycan-array binding and a solution-based fluorescence anisotropy assay. Each mutant was also tested in two biological assays to... (More)
Galectins are defined by a conserved beta-galactoside binding site, which has been linked to many of their important functions in e.g. cell adhesion, signaling and intracellular trafficking. Weak adjacent sites may enhance or decrease affinity for natural beta-galactoside containing glycoconjugates, but little is known about the biological role of this modulation of affinity (fine specificity). We have now produced 10 mutants of human galectin-3, with changes in these adjacent sites, which have altered carbohydrate-binding fine specificity but which retain the basic beta-galactoside binding activity as show by glycan-array binding and a solution-based fluorescence anisotropy assay. Each mutant was also tested in two biological assays to provide a correlation between fine specificity and function. Galectin-3 R186S, which has selectively lost affinity for LacNAc, a disaccharide moiety commonly found on glycoprotein glycans, has lost the ability to activate neutrophil leukocytes and intracellular targeting into vesicles. K176L has increased affinity for beta-galactosides substituted with GlcNAcbeta1-3 as found in poly-N-acetyllactosaminoglycans, and increased potency to activate neutrophil leukocytes, even though it has lost other aspects of galectin-3 fine specificity. G182A has altered carbohydrate-binding fine specificity and altered intracellular targeting into vesicles, a possible link to the intracellular galectin-3-mediated anti-apoptotic effect known to be lost by this mutant. Finally, the mutants have helped to define the differences in fine specificity shown by Xenopus, mouse and human galectin-3 and as such, evidence for adaptive change during evolution. (Less)
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Journal of Biological Chemistry
volume
285
issue
45
pages
35079 - 35091
publisher
ASBMB
external identifiers
  • wos:000283659100082
  • pmid:20807768
  • scopus:78049383095
ISSN
1083-351X
DOI
10.1074/jbc.M109.098160
language
English
LU publication?
yes
id
c59f8b52-53d9-415f-b35d-14fc021bbc68 (old id 1688659)
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http://www.ncbi.nlm.nih.gov/pubmed/20807768?dopt=Abstract
date added to LUP
2010-10-03 19:45:01
date last changed
2018-05-29 12:21:00
@article{c59f8b52-53d9-415f-b35d-14fc021bbc68,
  abstract     = {Galectins are defined by a conserved beta-galactoside binding site, which has been linked to many of their important functions in e.g. cell adhesion, signaling and intracellular trafficking. Weak adjacent sites may enhance or decrease affinity for natural beta-galactoside containing glycoconjugates, but little is known about the biological role of this modulation of affinity (fine specificity). We have now produced 10 mutants of human galectin-3, with changes in these adjacent sites, which have altered carbohydrate-binding fine specificity but which retain the basic beta-galactoside binding activity as show by glycan-array binding and a solution-based fluorescence anisotropy assay. Each mutant was also tested in two biological assays to provide a correlation between fine specificity and function. Galectin-3 R186S, which has selectively lost affinity for LacNAc, a disaccharide moiety commonly found on glycoprotein glycans, has lost the ability to activate neutrophil leukocytes and intracellular targeting into vesicles. K176L has increased affinity for beta-galactosides substituted with GlcNAcbeta1-3 as found in poly-N-acetyllactosaminoglycans, and increased potency to activate neutrophil leukocytes, even though it has lost other aspects of galectin-3 fine specificity. G182A has altered carbohydrate-binding fine specificity and altered intracellular targeting into vesicles, a possible link to the intracellular galectin-3-mediated anti-apoptotic effect known to be lost by this mutant. Finally, the mutants have helped to define the differences in fine specificity shown by Xenopus, mouse and human galectin-3 and as such, evidence for adaptive change during evolution.},
  author       = {Salomonsson, Emma and Carlsson, Michael and Osla, Veronica and Hendus-Altenburger, Ruth and Kahl Knutson, Barbro and Öberg, Christopher and Sundin, Anders and Nilsson, Rickard and Nordberg Karlsson, Eva and Nilsson, Ulf and Karlsson, Anna and Rini, James M and Leffler, Hakon},
  issn         = {1083-351X},
  language     = {eng},
  number       = {45},
  pages        = {35079--35091},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Mutational tuning of galectin-3 specificity and biological function.},
  url          = {http://dx.doi.org/10.1074/jbc.M109.098160},
  volume       = {285},
  year         = {2010},
}