Crystallization and preliminary X-ray diffraction studies of frutalin, an α-<scp>D</scp>-galactose-specific lectin from<i>Artocarpus incisa</i>seeds

Monteiro-Moreira, Ana Cristina de Oliveira; Pereira, H.M.; Neto, Antônio Eufrásio Vieira; Moreno, Frederico Bruno Mendes Batista; Lobo, Marina Duarte Pinto; Sousa, Felipe Domingos de; Moreira, Renato de Azevedo

doi:10.1107/s2053230x15015186

Cited by 4 publications

(4 citation statements)

References 27 publications

(25 reference statements)

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“…In SDS-PAGE, the profile of an isolated FTL batch matched the typical jacalin-like mass pattern of two bands ∼12 and 15 kDa, corresponding to monomers with different glycosylation levels, as previously reported [6]. Evidence based on FTL mass deconvoluted spectra suggests that the protein is naturally expressed as a mixture of isoforms [29]. Using a hemagglutination assay, we found FTL to be in a biologically active form and able to agglutinate human erythrocytes, which was preliminarily required before submitting the lectin to crystallization assays (Supplementary Figure S1).…”

Section: Resultssupporting

confidence: 81%

New structural insights into anomeric carbohydrate recognition by frutalin: an α-d-galactose-binding lectin from breadfruit seeds

Neto

Sousa

Pereira

et al. 2019

Biochemical Journal

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Frutalin (FTL) is a multiple-binding lectin belonging to the jacalin-related lectin (JRL) family and derived from Artocarpus incisa (breadfruit) seeds. This lectin specifically recognizes and binds α-d-galactose. FTL has been successfully used in immunobiological research for the recognition of cancer-associated oligosaccharides. However, the molecular bases by which FTL promotes these specific activities remain poorly understood. Here, we report the whole 3D structure of FTL for the first time, as determined by X-ray crystallography. The obtained crystals diffracted to 1.81 Å (Apo-frutalin) and 1.65 Å (frutalin–d-Gal complex) of resolution. The lectin exhibits post-translational cleavage yielding an α- (133 amino acids) and β-chain (20 amino acids), presenting a homotetramer when in solution, with a typical JRL β-prism. The β-prism was composed of three 4-stranded β-sheets forming three antiparallel Greek key motifs. The carbohydrate-binding site (CBS) involved the N-terminus of the α-chain and was formed by four key residues: Gly25, Tyr146, Trp147 and Asp149. Together, these results were used in molecular dynamics simulations in aqueous solutions to shed light on the molecular basis of FTL-ligand binding. The simulations suggest that Thr-Ser-Ser-Asn (TSSN) peptide excision reduces the rigidity of the FTL CBS, increasing the number of interactions with ligands and resulting in multiple-binding sites and anomeric recognition of α-d-galactose sugar moieties. Our findings provide a new perspective to further elucidate the versatility of FTL in many biological activities.

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Section: Resultssupporting

confidence: 81%

New structural insights into anomeric carbohydrate recognition by frutalin: an α-d-galactose-binding lectin from breadfruit seeds

Neto

Sousa

Pereira

et al. 2019

Biochemical Journal

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“…This gives Frutalin multiple-binding character, able to interact with different ligands using the same carbohydrate binding site. However, its highest affinity is for α-D-galactose, which we have shown to be anchored by a cavity of four residues (Gly25, Tyr146, Trp147 and Asp149) near the N-terminal of the α chain [35].…”

Section: Discussionmentioning

confidence: 85%

RETRACTED: Hydrogel and membrane scaffold formulations of Frutalin (breadfruit lectin) within a polysaccharide galactomannan matrix have potential for wound healing

Sousa

Vasconselos

Silva

et al. 2019

International Journal of Biological Macromolecules

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Plant lectins are carbohydrate-binding proteins, which can interact with cell surfaces to initiate both inflammatory and antiinflammatory pathways, as well as immunomodulatory functions. Here, we extracted, purified and part-characterized the bioactivity of four seed lectins, Jacalin, Frutalin, DAL and PNA, before evaluating their potential for wound healing in cultured human skin fibroblasts. Only Frutalin stimulated fibroblast migration in vitro, prompting further studies which established its low cytotoxicity (unaffected fibroblast viability at ≤1 mg/mL) and interaction with TLR-4 receptors. The lectin also increased pERK expression, a marker of fibroblast proliferation, and stimulated IL-6 secretion. The in vivo potential of Frutalin for wound healing was then assessed in hybrid combination with the polysaccharide galactomannan, purified from Caesalpinia pulcherrima seeds, using both hydrogel formulations and membrane scaffolds (lyophilized hydrogel). Physicalchemical characterization of the hybrid showed that lectin-galactomannan interactions increased the pseudoplastic behaviour of solutions, reducing viscosity and increasing Frutalin's concentration. Furthermore, infrared spectroscopy revealed-OH band displacement (from 3350 to 3595 cm-1), likely caused by interaction of Frutalin with galactose residues present on galactomannan chains, while average membrane porosity was 100 μm, sufficient to ensure water vapor permeability. The healing activities of each hybrid material at three different Frutalin to galactomannan ratios were then tested in a surgical mouse model of cutaneous excisional wound repair. Accelerated angiogenesis and increased fibroblast and keratinocyte proliferation were observed with the optimal hybrid recovering the lesioned area after 11 days. Our findings indicate Frutalin as a biomolecule with potential for tissue repair, regeneration and chronic wound healing.

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“…Among the lectins, frutalin (FTL) is extracted from the seeds of Artocarpus incisa L. This lectin has a tetrameric form and α-D-galactose specific link (Monteiro-Moreira et al 2015). Then, due to its binding characteristics, FTL can recognize and fix to cell membranes, and even penetrate into the cell nucleus, influencing cell-cell interactions, communication, proliferation, and cell death mechanism (Sharon 2004;Oliveira et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Structural characteristics and biotechnological applications of frutalin: lectin extracted from Artocarpus incisa

Silva

Aguiar

Silva

2018

CeN

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Frutalin is a lectin extracted from the seeds of Artocarpus incisa, which belongs to the Moreacae family. This family consists of approximately 75 genera and 1,550 tropical species that commonly found in pan-tropical regions. Frutalin has attracted the attention of researchers due to its ability to recognize carbohydrates in cell membranes with modifications in the glycosylation pattern. Therefore, frutalin presents a striking potential to be used as biomarker of cancer cells. Despite having a great biotechnological potential, research involving this lectin is still limited. Thus, the aim of this review is to discuss the structural and functional characteristics of frutalin, properties, the mechanisms of action, as well as the biotechnological applications of this lectin.

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Crystallization and preliminary X-ray diffraction studies of frutalin, an α-D-galactose-specific lectin fromArtocarpus incisaseeds

Cited by 4 publications

References 27 publications

New structural insights into anomeric carbohydrate recognition by frutalin: an α-d-galactose-binding lectin from breadfruit seeds

New structural insights into anomeric carbohydrate recognition by frutalin: an α-d-galactose-binding lectin from breadfruit seeds

RETRACTED: Hydrogel and membrane scaffold formulations of Frutalin (breadfruit lectin) within a polysaccharide galactomannan matrix have potential for wound healing

Structural characteristics and biotechnological applications of frutalin: lectin extracted from Artocarpus incisa

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