Isolation and biochemical characterization of a galactoside binding lectin from Bauhinia variegata candida (BvcL) seeds.
A new lectin (BvcL) from seeds of a primitive Brazilian Caesalpinoideae, the Bauhinia variegata candida was purified and biochemical characterized. BvcL was isolated by gel filtration chromatography on Sephadex G75 and affinity chromatography on immobilized D: -lactose column. SDS-PAGE showed that BvcL under non-reducing condition presents two bands of 68 and 32 kDa and a single band of 32 kDa in reducing condition. However, only one band was seen in native PAGE. The hemagglutination activity of BvcL was not specific for any human blood group trypsin-treated erythrocytes. Carbohydrate inhibition analysis indicated that BvcL is inhibited by lactose, galactose, galactosamine and other galactoside derivates. Amino acid analysis revealed a large content of Ser, Gly, Thr, Asp and Glu and low concentrations of Met, Cys and His. Intrinsic fluorescence of BvcL was not significantly affected by sugar binding galactose; and aromatic-region CD is unusually high for plant lectins. The N-terminal amino acid sequence of 17 residues showed 90% sequential homology to galactose-specific legume lectins of the subfamily Caesalpinoideae.
Silva
José A
JA
Department of Biochemistry, Institute of Biology, State University of Campinas (UNICAMP), 13083-970 Campinas, SP, Brazil.
Damico
Daniela C S
DC
Baldasso
Paulo A
PA
Mattioli
Marcelo A
MA
Winck
Flávia V
FV
Fraceto
Leonardo F
LF
Novello
José C
JC
Marangoni
Sérgio
S
D000595:Amino Acid Sequence; D000818:Animals; D036481:Bauhinia / Q000737:chemistry* / Q000145:classification; D015180:Electrophoresis, Gel, Two-Dimensional; D037161:Galectins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits; D012639:Seeds / Q000737:chemistry*; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D037161:Galectins; D037121:Plant Lectins