Purification, Biochemical Characterization, and Amino Acid Sequence of a Novel Type of Lectin from Aplysia dactylomela Eggs with Antibacterial/Antibiofilm Potential.
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 × 106M-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.
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
Rômulo Farias
RF
Carneiro
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
Renato Cézar Farias
RC
Torres
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
Renata Pinheiro
RP
Chaves
Laboratório Integrado de Biomoléculas - LIBS, Departamento de Patologia e Medicina Legal, Universidade Federal do Ceará, Monsenhor Furtado, s/n, Fortaleza, Ceará, 60430-160, Brazil.
Mayron Alves
MA
de Vasconcelos
Departamento de Física, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Fortaleza, Ceará, 60440-970, Brazil.
Bruno Lopes
BL
de Sousa
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
André Castelo Rodrigues
AC
Goveia
Laboratório Integrado de Biomoléculas - LIBS, Departamento de Patologia e Medicina Legal, Universidade Federal do Ceará, Monsenhor Furtado, s/n, Fortaleza, Ceará, 60430-160, Brazil.
Francisco Vassiliepe
FV
Arruda
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
Maria Nágila Carneiro
MN
Matos
Laboratório de Invertebrados Marinhos do Ceará - LIMCE, Departamento de Biologia, Universidade Federal do Ceará, Campus do Pici s/n, bloco 906, Fortaleza, CE, 60455-760, Brazil.
Helena
H
Matthews-Cascon
Departamento de Física, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Fortaleza, Ceará, 60440-970, Brazil.
Valder Nogueira
VN
Freire
Laboratório Integrado de Biomoléculas - LIBS, Departamento de Patologia e Medicina Legal, Universidade Federal do Ceará, Monsenhor Furtado, s/n, Fortaleza, Ceará, 60430-160, Brazil.
Edson Holanda
EH
Teixeira
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil.
Celso Shiniti
CS
Nagano
Laboratório de Biotecnologia Marinha - BioMar-Lab, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Campus do Pici s/n, bloco 871, Av. Mister Hull, Box 6043, Fortaleza, Ceará, 60440-970, Brazil. alexholandasampaio@gmail.com.
Alexandre Holanda
AH
Sampaio
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D001048:Aplysia / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005697:Galactosides / Q000494:pharmacology; D015870:Gene Expression; D006385:Hemagglutination Inhibition Tests; D006603:Hexuronic Acids / Q000494:pharmacology; D037102:Lectins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D062105:Molecular Docking Simulation; D000072417:Protein Domains; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000254:growth & development; D015053:Zygote / Q000737:chemistry*
D000900:Anti-Bacterial Agents; D005697:Galactosides; D006603:Hexuronic Acids; D037102:Lectins; D011994:Recombinant Proteins; C007819:galacturonic acid
Biofilm; Galacturonic acid; Lectin; Mass spectrometry; Sea hare