Mutational tuning of galectin-3 specificity and biological function.

Summary, in English

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: Emma Salomonsson; Michael Carlsson; Veronica Osla; Ruth Hendus-Altenburger; Barbro Kahl Knutson; Christopher Öberg; Anders Sundin; Rickard Nilsson; Eva Nordberg Karlsson; Ulf Nilsson; Anna Karlsson; James M Rini; Hakon Leffler
• Department/s: Division of Microbiology, Immunology and Glycobiology - MIG; Centre for Analysis and Synthesis; Biotechnology; Medical Radiation Physics, Malmö
• Publishing year: 2010
• Language: English
• Pages: 35079-35091
• Publication/Series: Journal of Biological Chemistry
• Volume: 285
• Issue: 45
• Links: http://www.ncbi.nlm.nih.gov/pubmed/20807768?dopt=Abstract; http://dx.doi.org/10.1074/jbc.M109.098160
• Document type: Journal article
• Publisher: American Society for Biochemistry and Molecular Biology
• Topic: Microbiology in the medical area; Radiology, Nuclear Medicine and Medical Imaging; Immunology in the medical area
• Status: Published
• Research group: Medical Radiation Physics, Malmö
• ISBN/ISSN/Other: ISSN: 1083-351X