Purification and molecular cloning of a new galactose-specific lectin from Bauhinia variegata seeds.
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.
Pinto
Luciano S
LS
Centro de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil. dmpluc@ufpel.edu.br
Nagano
Celso S
CS
Oliveira
Taianá M
TM
Moura
Tales R
TR
Sampaio
Alexandre H
AH
Debray
Henri
H
Pinto
Vicente P
VP
Dellagostin
Odir A
OA
Cavada
Benildo S
BS
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
D037121:Plant Lectins; D005690:Galactose