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Characterization of isoforms of the lectin isolated from the red algae Bryothamnion seaforthii and its pro-healing effect.

do Nascimento-Neto LG, Carneiro RF, da Silva SR, da Silva BR, Vassiliepe Sousa Arruda F, Carneiro VA, do Nascimento KS, Saker-Sampaio S, da Silva VA, Porto AL, Cavada BS, Sampaio AH, Teixeira EH, Nagano CS - Mar Drugs (2012)

Bottom Line: Because of their biotechnological potential, lectins are widely used in biomedical research.The lectin was purified using ion exchange chromatography with DEAE cellulose and characterized using tandem mass spectrometry.To evaluate the potential of each treatment, the animals were anesthetized and sacrificed on days 2, 7 and 12, respectively.

View Article: PubMed Central - PubMed

Affiliation: LIBS, Integrated Laboratory of Biomolecules, Faculty of Medicine of Sobral, Federal University of Ceará, Fortaleza, CE 60020-181, Brazil. ziullec@gmail.com

ABSTRACT
Lectins are a structurally heterogeneous group of proteins that have specific binding sites for carbohydrates and glycoconjugates. Because of their biotechnological potential, lectins are widely used in biomedical research. The present study aimed to evaluate the healing potential of the lectin isolated from the marine red alga Bryothamnion seaforthii (BSL). The lectin was purified using ion exchange chromatography with DEAE cellulose and characterized using tandem mass spectrometry. For healing tests, skin wounds were induced in the dorsal thoracic region of mice. These animals were randomly divided into three groups and subjected to topical treatment for 12 days with BSL, bovine serum albumin and 150 mM NaCl. To evaluate the potential of each treatment, the animals were anesthetized and sacrificed on days 2, 7 and 12, respectively. The parameters evaluated included the wound area, the proportion of wound closure and the histological diagnosis. The wound closure was more effective with BSL (Postoperative Day 7 and 12) than controls. The luminal epithelium was completely restructured; the presence of collagen in the dermis and the strongly active presence of young skin annexes demonstrate the potential of treatment with BSL compared with controls. Our findings suggest that BSL has pro-healing properties and can be a potential medical process in the treatment of acute wounds.

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Related in: MedlinePlus

MALDI-TOF-MS analysis of CA-BSL under denaturing and reducing conditions in the presence of iodoacetamide. The BSL samples treated with dithiothreitol (DTT) and iodoacetamide exhibited increased masses at 232 Daltons (4 × 58.00), suggesting the presence of four cysteine residues engaged in the formation of two intrachain disulfide bonds.
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marinedrugs-10-01936-f003: MALDI-TOF-MS analysis of CA-BSL under denaturing and reducing conditions in the presence of iodoacetamide. The BSL samples treated with dithiothreitol (DTT) and iodoacetamide exhibited increased masses at 232 Daltons (4 × 58.00), suggesting the presence of four cysteine residues engaged in the formation of two intrachain disulfide bonds.

Mentions: The existence of lectin isoforms was also observed in the lectins isolated from Bryothamnion triquetrum [19], Eucheuma serra [20] and Hypnea japonica [21]. The MALDI-TOF masses of intact BSL did not change upon incubation of the lectin with iodoacetamide under non-reducing conditions, indicating that BSL does not possess free sulfhydryl groups (data not shown). In contrast, a MALDI-TOF-MS analysis of denatured, reduced and carbamidomethylated BSL (CA-BSL) exhibited five major ions at m/z 9133.1 ± 2, 9194.1 ± 2, 9245.3 ± 2, 9296.5 ± 2 and 9328.7 ± 2, indicating the presence of four cysteine residues involved in two intrachain disulfide bonds (Figure 3).


Characterization of isoforms of the lectin isolated from the red algae Bryothamnion seaforthii and its pro-healing effect.

do Nascimento-Neto LG, Carneiro RF, da Silva SR, da Silva BR, Vassiliepe Sousa Arruda F, Carneiro VA, do Nascimento KS, Saker-Sampaio S, da Silva VA, Porto AL, Cavada BS, Sampaio AH, Teixeira EH, Nagano CS - Mar Drugs (2012)

MALDI-TOF-MS analysis of CA-BSL under denaturing and reducing conditions in the presence of iodoacetamide. The BSL samples treated with dithiothreitol (DTT) and iodoacetamide exhibited increased masses at 232 Daltons (4 × 58.00), suggesting the presence of four cysteine residues engaged in the formation of two intrachain disulfide bonds.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475265&req=5

marinedrugs-10-01936-f003: MALDI-TOF-MS analysis of CA-BSL under denaturing and reducing conditions in the presence of iodoacetamide. The BSL samples treated with dithiothreitol (DTT) and iodoacetamide exhibited increased masses at 232 Daltons (4 × 58.00), suggesting the presence of four cysteine residues engaged in the formation of two intrachain disulfide bonds.
Mentions: The existence of lectin isoforms was also observed in the lectins isolated from Bryothamnion triquetrum [19], Eucheuma serra [20] and Hypnea japonica [21]. The MALDI-TOF masses of intact BSL did not change upon incubation of the lectin with iodoacetamide under non-reducing conditions, indicating that BSL does not possess free sulfhydryl groups (data not shown). In contrast, a MALDI-TOF-MS analysis of denatured, reduced and carbamidomethylated BSL (CA-BSL) exhibited five major ions at m/z 9133.1 ± 2, 9194.1 ± 2, 9245.3 ± 2, 9296.5 ± 2 and 9328.7 ± 2, indicating the presence of four cysteine residues involved in two intrachain disulfide bonds (Figure 3).

Bottom Line: Because of their biotechnological potential, lectins are widely used in biomedical research.The lectin was purified using ion exchange chromatography with DEAE cellulose and characterized using tandem mass spectrometry.To evaluate the potential of each treatment, the animals were anesthetized and sacrificed on days 2, 7 and 12, respectively.

View Article: PubMed Central - PubMed

Affiliation: LIBS, Integrated Laboratory of Biomolecules, Faculty of Medicine of Sobral, Federal University of Ceará, Fortaleza, CE 60020-181, Brazil. ziullec@gmail.com

ABSTRACT
Lectins are a structurally heterogeneous group of proteins that have specific binding sites for carbohydrates and glycoconjugates. Because of their biotechnological potential, lectins are widely used in biomedical research. The present study aimed to evaluate the healing potential of the lectin isolated from the marine red alga Bryothamnion seaforthii (BSL). The lectin was purified using ion exchange chromatography with DEAE cellulose and characterized using tandem mass spectrometry. For healing tests, skin wounds were induced in the dorsal thoracic region of mice. These animals were randomly divided into three groups and subjected to topical treatment for 12 days with BSL, bovine serum albumin and 150 mM NaCl. To evaluate the potential of each treatment, the animals were anesthetized and sacrificed on days 2, 7 and 12, respectively. The parameters evaluated included the wound area, the proportion of wound closure and the histological diagnosis. The wound closure was more effective with BSL (Postoperative Day 7 and 12) than controls. The luminal epithelium was completely restructured; the presence of collagen in the dermis and the strongly active presence of young skin annexes demonstrate the potential of treatment with BSL compared with controls. Our findings suggest that BSL has pro-healing properties and can be a potential medical process in the treatment of acute wounds.

Show MeSH
Related in: MedlinePlus