title: 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.
authors.affiliation: Centro de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil. dmpluc@ufpel.edu.br
authors.forename: Luciano S
authors.initials: LS
authors.lastname: Pinto
authors.forename: Celso S
authors.initials: CS
authors.lastname: Nagano
authors.forename: Taianá M
authors.initials: TM
authors.lastname: Oliveira
authors.forename: Tales R
authors.initials: TR
authors.lastname: Moura
authors.forename: Alexandre H
authors.initials: AH
authors.lastname: Sampaio
authors.forename: Henri
authors.initials: H
authors.lastname: Debray
authors.forename: Vicente P
authors.initials: VP
authors.lastname: Pinto
authors.forename: Odir A
authors.initials: OA
authors.lastname: Dellagostin
authors.forename: Benildo S
authors.initials: BS
authors.lastname: Cavada
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D036481:Bauhinia / Q000737:chemistry* / Q000235:genetics; D005690:Galactose / Q000378:metabolism*; D006384:Hemagglutination; D006801:Humans; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits; D012639:Seeds / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D037121:Plant Lectins; D005690:Galactose