title: Mutational tuning of galectin-3 specificity and biological function.
abstract: 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.
authors.affiliation: Section MIG, Department of Laboratory Medicine, Lund University, 223 62 Lund, Sweden.
authors.forename: Emma
authors.initials: E
authors.lastname: Salomonsson
authors.forename: Michael C
authors.initials: MC
authors.lastname: Carlsson
authors.forename: Veronica
authors.initials: V
authors.lastname: Osla
authors.forename: Ruth
authors.initials: R
authors.lastname: Hendus-Altenburger
authors.forename: Barbro
authors.initials: B
authors.lastname: Kahl-Knutson
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Oberg
authors.forename: Anders
authors.initials: A
authors.lastname: Sundin
authors.forename: Rickard
authors.initials: R
authors.lastname: Nilsson
authors.forename: Eva
authors.initials: E
authors.lastname: Nordberg-Karlsson
authors.forename: Ulf J
authors.initials: UJ
authors.lastname: Nilsson
authors.forename: Anna
authors.initials: A
authors.lastname: Karlsson
authors.forename: James M
authors.initials: JM
authors.lastname: Rini
authors.forename: Hakon
authors.initials: H
authors.lastname: Leffler
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D005697:Galactosides / Q000235:genetics / Q000378:metabolism*; D037502:Galectin 3 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D051379:Mice; D020125:Mutation, Missense; D018375:Neutrophil Activation / Q000187:drug effects / Q000502:physiology*; D009504:Neutrophils / Q000378:metabolism*; D013379:Substrate Specificity; D014982:Xenopus laevis
chemical_list: D005697:Galactosides; D037502:Galectin 3; C485095:Lgals3 protein, mouse
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