Toward Comprehensive Analysis of the Galectin Network in Chicken: Unique Diversity of Galectin‐3 and Comparison of its Localization Profile in Organs of Adult Animals to the Other Four Members of this Lectin Family

Characterization of all members of a gene family established by gene divergence is essential to delineate distinct or overlapping expression profiles and functionalities. Their activity as potent modulators of diverse physiological processes directs interest to galectins (endogenous lectins with β‐sandwich fold binding β‐galactosides and peptide motifs), warranting their study with the long‐term aim of a comprehensive analysis. The comparatively low level of complexity of the galectin network in chicken with five members explains the choice of this organism as model. Previously, the three proto‐type chicken galectins CG‐1A, CG‐1B, and CG‐2 as well as the tandem‐repeat‐type CG‐8 had been analyzed. Our study fills the remaining gap to determine gene structure, protein characteristics and expression profile of the fifth protein, that is, chimera‐type chicken galectin‐3 (CG‐3). Its gene has a unique potential to generate variants: mRNA production stems from two promoters, alternative splicing of the form from the second transcription start point (tsp) can generate three mRNAs. The protein with functional phosphorylation sites in the N‐terminus generated by transcription from the first tsp (tsp1CG‐3) is the predominant CG‐3 type present in adult tissues. Binding assays with neoglycoproteins and cultured cells disclose marked similarity to properties of human galectin‐3. The expression and localization profiles as well as proximal promoter regions have characteristic features distinct from the other four CGs. This information on CG‐3 completes the description of the panel of CGs, hereby setting the stage for detailed comparative analysis of the entire CG family, e.g., in embryogenesis. Anat Rec, , 2011. © 2011 Wiley‐Liss, Inc.

[1]  H. Gabius,et al.  Towards Functional Glycomics by Lectin Histochemistry: Strategic Probe Selection to Monitor Core and Branch‐end Substitutions and Detection of Cell‐type and Regional Selectivity in Adult Mouse Testis and Epididymis , 2010, Anatomia, histologia, embryologia.

[2]  Christian Fischer,et al.  Tumor suppressor p16INK4a: Downregulation of galectin‐3, an endogenous competitor of the pro‐anoikis effector galectin‐1, in a pancreatic carcinoma model , 2010, The FEBS journal.

[3]  J. Kopitz,et al.  How adhesion/growth‐regulatory galectins‐1 and ‐3 attain cell specificity: Case study defining their target on neuroblastoma cells (SK‐N‐MC) and marked affinity regulation by affecting microdomain organization of the membrane , 2010, IUBMB life.

[4]  M. Kloor,et al.  N-domain of human adhesion/growth-regulatory galectin-9: preference for distinct conformers and non-sialylated N-glycans and detection of ligand-induced structural changes in crystal and solution. , 2010, The international journal of biochemistry & cell biology.

[5]  H. Gabius,et al.  Phosphorylation of adhesion- and growth-regulatory human galectin-3 leads to the induction of axonal branching by local membrane L1 and ERM redistribution , 2010, Journal of Cell Science.

[6]  N. Bovin,et al.  Carbamate-linked lactose: design of clusters and evidence for selectivity to block binding of human lectins to (neo)glycoproteins with increasing degree of branching and to tumor cells. , 2009, Bioconjugate chemistry.

[7]  W. Lehmann,et al.  Analysis of autophosphorylation sites in the recombinant catalytic subunit alpha of cAMP-dependent kinase by nano-UPLC–ESI–MS/MS , 2009, Analytical and bioanalytical chemistry.

[8]  Hans-Joachim Gabius,et al.  From structural to functional glycomics: core substitutions as molecular switches for shape and lectin affinity of N-glycans , 2009, Biological chemistry.

[9]  M. Boulton,et al.  Apoptosis gene profiling reveals spatio-temporal regulated expression of the p53/Mdm2 pathway during lens development , 2009, Experimental eye research.

[10]  H. Gabius,et al.  Unique chicken tandem-repeat-type galectin: implications of alternative splicing and a distinct expression profile compared to those of the three proto-type proteins. , 2009, Biochemistry.

[11]  W. Lehmann,et al.  Separation of peptide isomers and conformers by ultra performance liquid chromatography. , 2009, Journal of separation science.

[12]  H. Gabius,et al.  Homodimeric chicken galectin CG-1B (C-14): Crystal structure and detection of unique redox-dependent shape changes involving inter- and intrasubunit disulfide bridges by gel filtration, ultracentrifugation, site-directed mutagenesis, and peptide mass fingerprinting. , 2009, Journal of molecular biology.

[13]  H. Gabius,et al.  Phosphorylated Human Lectin Galectin‐3: Analysis of Ligand Binding by Histochemical Monitoring of Normal/Malignant Squamous Epithelia and by Isothermal Titration Calorimetry , 2009, Anatomia, histologia, embryologia.

[14]  H. Gabius,et al.  Cell-type-specific expression of murine multifunctional galectin-3 and its association with follicular atresia/luteolysis in contrast to pro-apoptotic galectins-1 and -7 , 2008, Histochemistry and Cell Biology.

[15]  W. Lehmann,et al.  Phosphorylated human galectin-3: facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy. , 2008, Biochimica et biophysica acta.

[16]  J. Kopitz,et al.  Prototype chicken galectins revisited: characterization of a third protein with distinctive hydrodynamic behaviour and expression pattern in organs of adult animals. , 2008, The Biochemical journal.

[17]  D. Kufe,et al.  The MUC1 and galectin-3 oncoproteins function in a microRNA-dependent regulatory loop. , 2007, Molecular cell.

[18]  Hiroaki Nakagawa,et al.  Tumor suppressor p16INK4a − modulator of glycomic profile and galectin‐1 expression to increase susceptibility to carbohydrate‐dependent induction of anoikis in pancreatic carcinoma cells , 2007, The FEBS journal.

[19]  A. Raz,et al.  The role of galectin-3 in cancer drug resistance. , 2007, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[20]  H. Gabius,et al.  Substitutions in the N-glycan core as regulators of biorecognition: the case of core-fucose and bisecting GlcNAc moieties. , 2007, Biochemistry.

[21]  Alexandre M J J Bonvin,et al.  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. , 2007, Glycobiology.

[22]  Zhichao Pei,et al.  Glycosyldisulfides from dynamic combinatorial libraries as O-glycoside mimetics for plant and endogenous lectins: their reactivities in solid-phase and cell assays and conformational analysis by molecular dynamics simulations. , 2006, Bioorganic & medicinal chemistry.

[23]  D. Kirsch,et al.  Effective solvation of alkaline earth ions by proline-rich proteolytic peptides of galectin-3 upon electrospray ionisation. , 2006, Rapid communications in mass spectrometry : RCM.

[24]  H. Gabius,et al.  Branching mode in complex-type triantennary N-glycans as regulatory element of their ligand properties. , 2006, Biochimica et Biophysica Acta.

[25]  H. Gabius,et al.  A Guide to Signaling Pathways Connecting Protein-Glycan Interaction with the Emerging Versatile Effector Functionality of Mammalian Lectins , 2006 .

[26]  H. Gabius,et al.  Galectins bind to the multivalent glycoprotein asialofetuin with enhanced affinities and a gradient of decreasing binding constants. , 2005, Biochemistry.

[27]  Zena Werb,et al.  Galectin-3 is a downstream regulator of matrix metalloproteinase-9 function during endochondral bone formation. , 2005, Molecular biology of the cell.

[28]  Hans-Joachim Gabius,et al.  Growth-regulatory human galectin-1: crystallographic characterisation of the structural changes induced by single-site mutations and their impact on the thermodynamics of ligand binding. , 2004, Journal of molecular biology.

[29]  S. Sidhu,et al.  Phylogenetic analysis of the vertebrate galectin family. , 2004, Molecular biology and evolution.

[30]  H. Gabius,et al.  Galectin-3 Precipitates as a Pentamer with Synthetic Multivalent Carbohydrates and Forms Heterogeneous Cross-linked Complexes* , 2004, Journal of Biological Chemistry.

[31]  Fu-Tong Liu,et al.  New aspects of galectin functionality in nuclei of cultured bone marrow stromal and epidermal cells: biotinylated galectins as tool to detect specific binding sites , 2003, Biology of the cell.

[32]  Douglas N W Cooper,et al.  Galectinomics: finding themes in complexity. , 2002, Biochimica et biophysica acta.

[33]  Fu-Tong Liu,et al.  Thermodynamic binding studies of cell surface carbohydrate epitopes to galectins-1, -3, and -7: Evidence for differential binding specificities , 2002 .

[34]  A. Artigues,et al.  New Alternatively Spliced Form of Galectin-3, a Member of the β-Galactoside-binding Animal Lectin Family, Contains a Predicted Transmembrane-spanning Domain and a Leucine Zipper Motif* , 2002, The Journal of Biological Chemistry.

[35]  Klaus Kayser,et al.  Structure-activity profiles of complex biantennary glycans with core fucosylation and with/without additional alpha 2,3/alpha 2,6 sialylation: synthesis of neoglycoproteins and their properties in lectin assays, cell binding, and organ uptake. , 2002, Journal of medicinal chemistry.

[36]  H Kaltner,et al.  Negative Regulation of Neuroblastoma Cell Growth by Carbohydrate-dependent Surface Binding of Galectin-1 and Functional Divergence from Galectin-3* , 2001, The Journal of Biological Chemistry.

[37]  A. M. Wu,et al.  Carbohydrate specificity of a galectin from chicken liver (CG-16). , 2001, The Biochemical journal.

[38]  E. Boja,et al.  Overalkylation of a protein digest with iodoacetamide. , 2001, Analytical chemistry.

[39]  A. Surolia,et al.  Thermodynamic analysis of the binding of galactose and poly‐N‐acetyllactosamine derivatives to human galectin‐3 , 2001, FEBS letters.

[40]  A. Legrand,et al.  Identification of an Internal Gene to the Human Galectin-3 Gene with Two Different Overlapping Reading Frames That Do Not Encode Galectin-3* , 2001, The Journal of Biological Chemistry.

[41]  H. Gabius,et al.  Temporal and Spatial Regulation of Expression of two Galectins During Kidney Development of the Chicken , 2000, The Histochemical Journal.

[42]  H. Gabius,et al.  The 2.15 A crystal structure of CG-16, the developmentally regulated homodimeric chicken galectin. , 1999, Journal of molecular biology.

[43]  M. Monsigny,et al.  Modulation of the expression of the rabbit galectin-3 gene by p53 and c-Ha-ras proteins and PMA. , 1997, Glycobiology.

[44]  H Kaltner,et al.  Involvement of laser photo-CIDNP (chemically induced dynamic nuclear polarization)-reactive amino acid side chains in ligand binding by galactoside-specific lectins in solution. , 1997, European journal of biochemistry.

[45]  T. Linsenmayer,et al.  Identification and characterization of up‐regulated genes during chondrocyte hypertrophy , 1996, Developmental dynamics : an official publication of the American Association of Anatomists.

[46]  H Kaltner,et al.  Different Architecture of the Combining Site of the Two Chicken Galectins Revealed by Chemical Mapping Studies with Synthetic Ligand Derivatives* , 1996, The Journal of Biological Chemistry.

[47]  F. Zindy,et al.  Alternative reading frames of the INK4a tumor suppressor gene encode two unrelated proteins capable of inducing cell cycle arrest , 1995, Cell.

[48]  M. Monsigny,et al.  The second intron of the human galectin‐3 gene has a strong promoter activity down‐regulated by p53 , 1995, FEBS letters.

[49]  John L. Wang,et al.  Galectin-3: differential accumulation of distinct mRNAs in serum-stimulated mouse 3T3 fibroblasts , 1994, Glycoconjugate Journal.

[50]  R C Hughes,et al.  Mac-2: a versatile galactose-binding protein of mammalian tissues. , 1994, Glycobiology.

[51]  S. Spicer,et al.  Diversity of cell glycoconjugates shown histochemically: a perspective. , 1992, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[52]  J. Hirabayashi,et al.  Structure of chicken 16-kDa beta-galactoside-binding lectin. Complete amino acid sequence, cloning of cDNA, and production of recombinant lectin. , 1990, The Journal of biological chemistry.

[53]  U. Brinck,et al.  Binding of T-antigen-bearing neoglycoprotein and peanut agglutinin to cultured tumor cells and breast carcinomas. , 1990, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[54]  S. Weiner,et al.  The Mac-2 antigen is a galactose-specific lectin that binds IgE , 1989, The Journal of experimental medicine.

[55]  A. Schauer,et al.  Endogenous sugar‐binding proteins in human breast tissue and benign and malignant breast lesions , 1988, Cancer.

[56]  J. L. Wang,et al.  Carbohydrate-binding protein 35: identification of the galactose-specific lectin in various tissues of mice , 1984, Molecular and cellular biology.

[57]  S. Barondes,et al.  Soluble lectins: a new class of extracellular proteins. , 1984, Science.

[58]  K. Kasai,et al.  Purification and characterization of beta-galactoside-binding lectin from chick embryonic skin. , 1983, Biochimica et biophysica acta.

[59]  T. Flotte,et al.  Dendritic cell and macrophage staining by monoclonal antibodies in tissue sections and epidermal sheets. , 1983, The American journal of pathology.

[60]  S. Barondes,et al.  Two lactose binding lectins from chicken tissues. Purified lectin from intestine is different from those in liver and muscle. , 1980, The Journal of biological chemistry.

[61]  H. Gabius.  The sugar code : fundamentals of glycosciences , 2009 .

[62]  P. Stanley,et al.  Lectin-resistant CHO glycosylation mutants. , 2006, Methods in enzymology.

[63]  M. Vidal,et al.  Unique sequence and expression profiles of rat galectins-5 and -9 as a result of species-specific gene divergence. , 2006, The international journal of biochemistry & cell biology.

[64]  Hans-Joachim Gabius,et al.  Cell surface glycans: the why and how of their functionality as biochemical signals in lectin-mediated information transfer. , 2006, Critical reviews in immunology.

[65]  H. Gabius,et al.  Lectin localization in human nerve by biochemically defined lectin-binding glycoproteins, neoglycoprotein and lectin-specific antibody , 2004, Histochemistry.

[66]  K. Kayser,et al.  Determination of modulation of ligand properties of synthetic complex-type biantennary N-glycans by introduction of bisecting GlcNAc in silico, in vitro and in vivo. , 2004, European journal of biochemistry.

[67]  John L. Wang,et al.  The human LGALS3 (galectin-3) gene: determination of the gene structure and functional characterization of the promoter. , 1998, Archives of biochemistry and biophysics.

[68]  J. Hirabayashi,et al.  Galectins: a family of animal lectins that decipher glycocodes. , 1996, Journal of biochemistry.

[69]  H. Gabius,et al.  Contribution of carbohydrate histochemistry to glycobiology. , 1994, Histology and histopathology.

[70]  H. Gabius.  Endogenous lectins in tumors and the immune system. , 1987, Cancer investigation.