title: Activity-structure correlations in divergent lectin evolution: fine specificity of chicken galectin CG-14 and computational analysis of flexible ligand docking for CG-14 and the closely related CG-16.
abstract: Gene duplication and sequence divergence are driving forces toward establishing protein families. To examine how sequence changes affect carbohydrate specificity, the two closely related proto-type chicken galectins CG-14 and CG-16 were selected as models. Binding properties were analyzed using a highly sensitive solid-phase assay. We tested 56 free saccharides and 34 well-defined glycoproteins. The two galectins share preference for the II (Galbeta1-4GlcNAc) versus I (Galbeta1-3GlcNAc) version of beta-galactosides. A pronounced difference is found owing to the reactivity of CG-14 with histo-blood group ABH active oligosaccharides and A/B active glycoproteins. These experimental results prompted to determine activity-structure correlations by modeling. Computational analysis included consideration of the flexibility of binding partners and the presence of water molecules. It provided a comparative description of complete carbohydrate recognition domains, which had so far not been characterized in animal galectins. The structural models assigned II, I selectivity to a region downstream of the central Trp moiety. Docking revealed that the tetrasaccharides can be accommodated in their free-state low-energy conformations. CG-14's preference for A versus B epitopes could be attributed to a contact between His124 and the N-acetyl group of GalNAc. Regarding intergalectin comparison, the Ala53/Cys51 exchange affects the interaction potential of His54/His52. Close inspection of simulated dynamic interplay revealed reorientation of His124 at the site of the His124/Glu123 substitution, with potential impact on ligand dissociation. In summary, this study identifies activity differences and provides information on their relation to structural divergence, epitomizing the value of this combined approach beyond galectins.
authors.affiliation: Glyco-Immunochemistry Research Laboratory, Institute of Molecular and Cellular Biology, College of Medicine, Chang-Gung University, Kwei-San, Tao-Yuan, Taiwan. amwu@mail.cgu.edu.tw
authors.forename: Albert M
authors.initials: AM
authors.lastname: Wu
authors.forename: Tanuja
authors.initials: T
authors.lastname: Singh
authors.forename: Jia-Hau
authors.initials: JH
authors.lastname: Liu
authors.forename: Mickael
authors.initials: M
authors.lastname: Krzeminski
authors.forename: Roland
authors.initials: R
authors.lastname: Russwurm
authors.forename: Hans-Christian
authors.initials: HC
authors.lastname: Siebert
authors.forename: Alexandre M J J
authors.initials: AM
authors.lastname: Bonvin
authors.forename: Sabine
authors.initials: S
authors.lastname: André
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Gabius
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism; D002645:Chickens; D019295:Computational Biology; D019143:Evolution, Molecular; D005697:Galactosides / Q000378:metabolism*; D037161:Galectins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D020440:Gene Duplication; D006801:Humans; D037102:Lectins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D008954:Models, Biological; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000494:pharmacology; D011487:Protein Conformation; D013329:Structure-Activity Relationship
chemical_list: D015295:Antigens, Tumor-Associated, Carbohydrate; D005697:Galactosides; D037161:Galectins; D037102:Lectins; D008024:Ligands; D009844:Oligosaccharides; C016467:Thomsen-Friedenreich antigen