Purification and molecular cloning of a new galactose-specific lectin from Bauhinia variegata seeds

Abstract

A new galactose-specific lectin was purified from seeds of a Caesalpinoideae plant, Bauhinia variegata, by affinity chromatography on lactose-agarose. Protein extracts haemagglutinated rabbit and human erythrocytes (native and treated with proteolytic enzymes), showing preference for rabbit blood treated with papain and trypsin. Among various carbohydrates tested, the lectin was best inhibited by D-galactose and its derivatives, especially lactose. SDS-PAGE showed that the lectin, named BVL, has a pattern similar to other lectins isolated from the same genus, Bauhinia purpurea agglutinin (BPA). The molecular mass of BVL subunit is 32 871 Da, determined by MALDI-TOF spectrometry. DNA extracted from B.variegata young leaves and primers designed according to the B. purpurea lectin were used to generate specific fragments which were cloned and sequenced, revealing two distinct isoforms. The bvl gene sequence comprised an open reading frame of 876 base pairs which encodes a protein of 291 amino acids. The protein carried a putative signal peptide. The mature protein was predicted to have 263 amino acid residues and 28 963 Da in size.

Antibacterial and Hemolytic Activity of a new Lectin Purified from the Seeds of Sterculia Foetida L.

“…These enzymes act thru cleavage of certain membrane proteins, further exposing membrane carbohydrates comprising the glycocalyx of the erythrocyte membrane. This allows the lectin more access to surface carbohydrates, which might increase the hemagglutination titers [24,25].…”

Investigation of cellular proliferative potential of lectin extracted from Bauhinia variegata according to different cell lines

“…This lectin, when used in skin wounds in mice, was responsible for the epithelium reconstruction and increased keratin deposition, indicating improved wound healing [4]. Partial results of this experiment showed no cytotoxicity for the lineages tested, when compared to the control groups.…”

“…The native lectin B. variegata (BVL) was extracted from seeds, according to the protocol described by Pinto et al [4]. The protein was characterized, quantified and tested for hemagglutination activity.…”

The galactose‐binding lectin isolated from Bauhinia bauhinioides Mart seeds inhibits neutrophil rolling and adhesion via primary cytokines

“…Sequence heterogeneity was observed in one peptide D/S (T2/T2'), indicating that BBL preparations contain highly homologous isolectins. Similarly, the cloning and sequence of the B. variegata lectin gene have suggested that this protein could be encoded by a family of genes, which led to the identification of two isoforms (Pinto et al, 2008).…”