Purification, Biochemical Characterization, and Amino Acid Sequence of a Novel Type of Lectin from Aplysia dactylomela Eggs with Antibacterial/Antibiofilm Potential
Autor: | Mayron Alves de Vasconcelos, André Castelo Rodrigues Goveia, Renato Cézar Farias Torres, Rômulo Farias Carneiro, Alexandre Holanda Sampaio, Edson Holanda Teixeira, Helena Matthews-Cascon, Renata Pinheiro Chaves, Francisco Vassiliepe Sousa Arruda, Maria Nágila Carneiro Matos, Celso Shiniti Nagano, Bruno Lopes de Sousa, Valder N. Freire |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Staphylococcus aureus Glycosylation Zygote Sea hare Gene Expression Mannose Applied Microbiology and Biotechnology Protein Structure Secondary 03 medical and health sciences chemistry.chemical_compound Protein Domains Affinity chromatography Lectins Aplysia Escherichia coli Animals Amino Acid Sequence Peptide sequence Mass spectrometry Molecular mass biology Hexuronic Acids Biofilm Protein primary structure Lectin Galactosides Hemagglutination Inhibition Tests Recombinant Proteins Protein tertiary structure Anti-Bacterial Agents Molecular Docking Simulation 030104 developmental biology chemistry Biochemistry Biofilms Galacturonic acid biology.protein Sequence Alignment |
Zdroj: | Repositório Institucional da Universidade Federal do Ceará (UFC) Universidade Federal do Ceará (UFC) instacron:UFC |
ISSN: | 1436-2236 1436-2228 |
Popis: | A new lectin from Aplysia dactylomela eggs (ADEL) was isolated by affinity chromatography on HCl-activated Sepharose™ media. Hemagglutination caused by ADEL was inhibited by several galactosides, mainly galacturonic acid (Ka = 6.05 × 106 M−1). The primary structure of ADEL consists of 217 residues, including 11 half-cystines involved in five intrachain and one interchain disulfide bond, resulting in a molecular mass of 57,228 ± 2 Da, as determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry. ADEL showed high similarity with lectins isolated from Aplysia eggs, but not with other known lectins, indicating that these lectins could be grouped into a new family of animal lectins. Three glycosylation sites were found in its polypeptide backbone. Data from peptide-N-glycosidase F digestion and MS suggest that all oligosaccharides attached to ADEL are high in mannose. The secondary structure of ADEL is predominantly β-sheet, and its tertiary structure is sensitive to the presence of ligands, as observed by CD. A 3D structure model of ADEL was created and shows two domains connected by a short loop. Domain A is composed of a flat three-stranded and a curved five-stranded β-sheet, while domain B presents a flat three-stranded and a curved four-stranded β-sheet. Molecular docking revealed favorable binding energies for interactions between lectin and galacturonic acid, lactose, galactosamine, and galactose. Moreover, ADEL was able to agglutinate and inhibit biofilm formation of Staphylococcus aureus, suggesting that this lectin may be a potential alternative to conventional use of antimicrobial agents in the treatment of infections caused by Staphylococcal biofilms. |
Databáze: | OpenAIRE |
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