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Mutational Tuning of Galectin-3 Specificity and Biological Function
Galectins are defined by a conserved β-galactoside binding site that 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 β-galactoside-containing glycoconjugates, but li...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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American Society for Biochemistry and Molecular Biology
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2966122/ https://www.ncbi.nlm.nih.gov/pubmed/20807768 http://dx.doi.org/10.1074/jbc.M109.098160 |
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author | Salomonsson, Emma Carlsson, Michael C. Osla, Veronica Hendus-Altenburger, Ruth Kahl-Knutson, Barbro Öberg, Christopher T. Sundin, Anders Nilsson, Rickard Nordberg-Karlsson, Eva Nilsson, Ulf J. Karlsson, Anna Rini, James M. Leffler, Hakon |
author_facet | Salomonsson, Emma Carlsson, Michael C. Osla, Veronica Hendus-Altenburger, Ruth Kahl-Knutson, Barbro Öberg, Christopher T. Sundin, Anders Nilsson, Rickard Nordberg-Karlsson, Eva Nilsson, Ulf J. Karlsson, Anna Rini, James M. Leffler, Hakon |
author_sort | Salomonsson, Emma |
collection | PubMed |
description | Galectins are defined by a conserved β-galactoside binding site that 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 β-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 that have altered carbohydrate-binding fine specificity but that retain the basic β-galactoside binding activity as shown 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 β-galactosides substituted with GlcNAcβ1–3, 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, the evidence for adaptive change during evolution. |
format | Text |
id | pubmed-2966122 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-29661222011-01-04 Mutational Tuning of Galectin-3 Specificity and Biological Function Salomonsson, Emma Carlsson, Michael C. Osla, Veronica Hendus-Altenburger, Ruth Kahl-Knutson, Barbro Öberg, Christopher T. Sundin, Anders Nilsson, Rickard Nordberg-Karlsson, Eva Nilsson, Ulf J. Karlsson, Anna Rini, James M. Leffler, Hakon J Biol Chem Glycobiology and Extracellular Matrices Galectins are defined by a conserved β-galactoside binding site that 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 β-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 that have altered carbohydrate-binding fine specificity but that retain the basic β-galactoside binding activity as shown 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 β-galactosides substituted with GlcNAcβ1–3, 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, the evidence for adaptive change during evolution. American Society for Biochemistry and Molecular Biology 2010-11-05 2010-08-31 /pmc/articles/PMC2966122/ /pubmed/20807768 http://dx.doi.org/10.1074/jbc.M109.098160 Text en © 2010 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles |
spellingShingle | Glycobiology and Extracellular Matrices Salomonsson, Emma Carlsson, Michael C. Osla, Veronica Hendus-Altenburger, Ruth Kahl-Knutson, Barbro Öberg, Christopher T. Sundin, Anders Nilsson, Rickard Nordberg-Karlsson, Eva Nilsson, Ulf J. Karlsson, Anna Rini, James M. Leffler, Hakon Mutational Tuning of Galectin-3 Specificity and Biological Function |
title | Mutational Tuning of Galectin-3 Specificity and Biological Function |
title_full | Mutational Tuning of Galectin-3 Specificity and Biological Function |
title_fullStr | Mutational Tuning of Galectin-3 Specificity and Biological Function |
title_full_unstemmed | Mutational Tuning of Galectin-3 Specificity and Biological Function |
title_short | Mutational Tuning of Galectin-3 Specificity and Biological Function |
title_sort | mutational tuning of galectin-3 specificity and biological function |
topic | Glycobiology and Extracellular Matrices |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2966122/ https://www.ncbi.nlm.nih.gov/pubmed/20807768 http://dx.doi.org/10.1074/jbc.M109.098160 |
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