Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth

Autor: Jacek Lubkowski, David Farnsworth, Mariana Cristina Cabral Silva, Alexander Wlodawer, Sarah V. Durbin, Maria Luiza Vilela Oliva, Jeffrey C. Gildersleeve
Rok vydání: 2017
Předmět:
Models
Molecular
0301 basic medicine
Acetylgalactosamine
Tn antigen
Protein Data Bank (RCSB PDB)
Gene Expression
Oligosaccharides
Peptide
Crystallography
X-Ray
01 natural sciences
Biochemistry
Protein Structure
Secondary
Substrate Specificity
Cloning
Molecular
carbohydrate binding
chemistry.chemical_classification
Globosides
cancer cell growth inhibition
Glycopeptides
Recombinant Proteins
Bauhinia
Blood Group Antigens
Plant Lectins
Dimerization
Protein Binding
crystal structure
Glycan
Biology
010402 general chemistry
Article
03 medical and health sciences
Molecular recognition
Antigen
Cell Line
Tumor
Escherichia coli
Humans
Antigens
Tumor-Associated
Carbohydrate
Protein Interaction Domains and Motifs
Molecular Biology
Binding Sites
Plant Extracts
Cell growth
Lectin
Hydrogen Bonding
Cell Biology
Antineoplastic Agents
Phytogenic
0104 chemical sciences
Kinetics
030104 developmental biology
chemistry
biology.protein
lectin
Zdroj: Repositório Institucional da UNIFESP
Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
ISSN: 1742-4658
1742-464X
Popis: National Institutes of Health (NIH), National Cancer Institute, Center for Cancer Research Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) FAPESP (PD-BEPE) U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences Lectins have been used at length for basic research and clinical applications. New insights into the molecular recognition properties enhance our basic understanding of carbohydrate-protein interactions and aid in the design/development of new lectins. In this study, we used a combination of cell-based assays, glycan microarrays, and X-ray crystallography to evaluate the structure and function of the recombinant Bauhinia forficata lectin (BfL). The lectin was shown to be cytostatic for several cancer cell lines included in the NCI-60 panel in particular, it inhibited growth of melanoma cancer cells (LOX IMVI) by over 95%. BfL is dimeric in solution and highly specific for binding of oligosaccharides and glycopeptides with terminal N-acetylgalactosamine (GalNAc). BfL was found to have especially strong binding (apparent K-d = 0.5-1.0 nM) to the tumor-associated Tn antigen. High-resolution crystal structures were determined for the ligand-free lectin, as well as for its complexes with three Tn glycopeptides, globotetraose, and the blood group A antigen. Extensive analysis of the eight crystal structures and comparison to structures of related lectins revealed several unique features of GalNAc recognition. Of special note, the carboxylate group of Glu126, lining the glycan-binding pocket, forms H-bonds with both the N-acetyl of GalNAc and the peptide amido group of Tn antigens. Stabilization provided by Glu126 is described here for the first time for any GalNAc-specific lectin. Taken together, the results provide new insights into the molecular recognition of carbohydrates and provide a structural understanding that will enable rational engineering of BfL for a variety of applications. Database Structural data are available in the PDB under the accession numbers 5T50, 5T52, 5T55, 5T54, 5T5L, 5T5J, 5T5P, and 5T5O. NCI, Macromol Crystallog Lab, Ctr Canc Res, Frederick, MD 21702 USA NCI, Biol Chem Lab, Ctr Canc Res, Frederick, MD 21701 USA Univ Fed Sao Paulo, Escola Paulista Med, Sao Paulo, SP, Brazil Univ Fed Sao Paulo, Escola Paulista Med, Sao Paulo, SP, Brazil FAPESP: 2009/53766-5 FAPESP: 2012/06366-4 FAPESP: 2014/22649-1 FAPESP (PD-BEPE): 2014/22649-1 U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences: W-31-109-Eng-38 Web of Science
Databáze: OpenAIRE
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