Journal ArticleDOI
Engineering of a 3'-sulpho-Galβ1-4GlcNAc-specific probe by a single amino acid substitution of a fungal galectin.
Dan Hu,Hang Huang,Hiroaki Tateno,Shin-ichi Nakakita,Takashi Sato,Hisashi Narimatsu,Xinsheng Yao,Jun Hirabayashi +7 more
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TLDR
In the course of molecular engineering of a fungal galectin from Agrocybe cylindracea, a single substitution of Glu86 with alanine resulted in acquisition of specific binding for the 3'-sulpho-Galβ1-4GlcNAc structure, and extensive glyco-technological analysis revealed that this property was obtained in a 'loss of function' manner.Abstract:
Among sulphated glycans, little is known about 3'-sulphation because of the lack of useful probes. In the course of molecular engineering of a fungal galectin from Agrocybe cylindracea, we found that a single substitution of Glu86 with alanine resulted in acquisition of specific binding for the 3'-sulpho-Galβ1-4GlcNAc structure. Extensive glyco-technological analysis revealed that this property was obtained in a 'loss-of-function' manner. Though this mutant (E86A) had low total affinity, it showed substantial binding to a naturally occurring N-glycan, of which the terminal galactose is 3-sulphated. Moreover, E86A specifically bound to HeLa cells, in which galactose-3-O-sulfotransferases (Gal3ST2 or Gal3ST3) were over-expressed.read more
Citations
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Novel sulfated ligands for the cell adhesion molecule E-selectin rreveled by the neoglycolipid technology among O-liked oligo-saccharides on an ovarin cystadenoma glycoprotein
TL;DR: To the authors' knowledge this is the first report of a sulfofucooligosaccharide ligand for E-selectin, and the binding activity is substantially greater than those of lipid-linked Le(a) and Le(x)/SSEA-1 sequences and is at least equal to that of the 3'-sialyl-Le(x))/S SEA-1 glycolipid analogue.
Journal ArticleDOI
Lectin Engineering, a Molecular Evolutionary Approach to Expanding the Lectin Utilities
TL;DR: The current state of the art of lectin engineering is summarized and recent technological advances in this field including selection of template lectin, construction of a mutagenesis library, and high-throughput screening methods are discussed.
Journal ArticleDOI
Strategies and Tactics for the Development of Selective Glycan-Binding Proteins.
TL;DR: Glycan-binding proteins (GBPs) as mentioned in this paper are a powerful tool for interrogating glycan structure and function because of their ability to selectively bind defined carbohydrate epitopes and glycosidic linkages.
Journal ArticleDOI
Studying the Structural Significance of Galectin Design by Playing a Modular Puzzle: Homodimer Generation from Human Tandem-Repeat-Type (Heterodimeric) Galectin-8 by Domain Shuffling
Anna-Kristin Ludwig,Malwina Michalak,Nadya Shilova,Sabine André,Herbert Kaltner,Nicolai V. Bovin,Jürgen Kopitz,Hans-Joachim Gabius +7 more
TL;DR: Engineering of variants by changing the topological display of carbohydrate recognition domains (CRDs) provides tools to understand the inherent specificity of the functional pairing, and substantiates the potential for comparative functional studies between the variant and natural Gal-8 (NC)/Gal-8N.
Posted ContentDOI
Selectivity and engineering of the sialoglycan-binding spectrum in Siglec-like adhesins
Barbara A. Bensing,Lioudmila V. Loukachevitch,Rupesh Agarwal,Rupesh Agarwal,Izumi Yamakawa,Kelvin Luong,Azadeh Hadadianpour,Hai Yu,Kevin P. Fialkowski,Kevin P. Fialkowski,Manuel Castro,Zdzislaw Wawrzak,Xi Chen,Jerome Baudry,Jeremy C. Smith,Paul M. Sullam,Tina M. Iverson +16 more
TL;DR: The molecular basis for receptor selectivity in streptococci that bind to sialoglycan receptors is identified and how to convert these adhesins into selective probes that measure triand tetrasacharides within the context of larger glycosylations is revealed.
References
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Galectins. Structure and function of a large family of animal lectins.
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Introduction to galectins
TL;DR: A brief introduction to the galectins as a protein family with some comments on nomenclature is given.
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Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling.
Atsushi Kuno,Noboru Uchiyama,Shiori Koseki-Kuno,Youji Ebe,Seigo Takashima,Masao Yamada,Jun Hirabayashi +6 more
TL;DR: A new microarray procedure based on an evanescent-field fluorescence-detection principle that allows sensitive, real-time observation of multiple lectin-carbohydrate interactions under equilibrium conditions is described, which should contribute to creation of a new paradigm for glycomics.
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Glycosaminoglycans: key players in cancer cell biology and treatment
Nikos Afratis,Chrisostomi Gialeli,Dragana Nikitovic,Theodore Tsegenidis,Evgenia Karousou,Achilleas D. Theocharis,Mauro S. G. Pavão,George N. Tzanakakis,Nikos K. Karamanos +8 more
TL;DR: How the fine structural characteristics of glycosaminoglycans, and enzymes involved in their biosynthesis and degradation, are involved in cell signaling, cell function and cancer progression is summarized.
Journal ArticleDOI
Glycoprofiling with micro-arrays of glycoconjugates and lectins
S. Angeloni,J. L. Ridet,N. Kusy,H. Gao,F. Crevoisier,S. Guinchard,S. Kochhar,Hans Sigrist,Norbert Sprenger +8 more
TL;DR: Th thin film-coated photoactivatable surfaces applied for covalent immobilization of glycans, glycoconjugates, or lectins in microarray formats demonstrate the feasibility and versatility of the novel platforms for glycan profiling.