Purification, characterization and partial sequence of a pro-inflammatory lectin from seeds of Canavalia oxyphylla Standl. & L. O. Williams.
Recent studies have shown that lectins are promising tools for use in various biotechnological processes, as well as studies of various pathological mechanisms, isolation, and characterization of glycoconjugates and understanding the mechanisms underlying pathological mechanisms conditions, including the inflammatory response. This study aimed to purify, characterize physicochemically, and predict the biological activity of Canavalia oxyphylla lectin (CoxyL) in vitro and in vivo. CoxyL was purified by a single-step affinity chromatography in Sephadex® G-50 column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that the pure lectin consists of a major band of 30 kDa (α-chain) and two minor components (β-chain and γ-chain) of 16 and 13 kDa, respectively. These data were further confirmed by electrospray ionization mass spectrometry, suggesting that CoxyL is a typical ConA-like lectin. In comparison with the average molecular mass of α-chain, the partial amino acid sequence obtained corresponds to approximately 45% of the total CoxyL sequence. CoxyL presented hemagglutinating activity that was specifically inhibited by monosaccharides (D-glucose, D-mannose, and α-methyl-D-mannoside) and glycoproteins (ovalbumin and fetuin). Moreover, CoxyL was shown to be thermostable, exhibiting full hemagglutinating activity up to 60°C, and it was pH-sensitive for 1 h, exhibiting maximal activity at pH 7.0. CoxyL caused toxicity to Artemia nauplii and induced paw edema in rats. This biological activity highlights the importance of lectins as important tools to better understand the mechanisms underlying inflammatory responses.
Laboratório de Moléculas Biologicamente Ativas (Biomol-Lab), Department of Biochemistry and Molecular Biology, Federal University of Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza, CE, 60440-970, Brazil.
Mayara Q
MQ
Santiago
Cintia C F
CC
Leitão
Francisco N
FN
Pereira
Vanir R
VR
Pinto
Vinicius J S
VJ
Osterne
Claudia F
CF
Lossio
João B
JB
Cajazeiras
Helton C
HC
Silva
Francisco V S
FV
Arruda
Livia P
LP
Pereira
Ana M S
AM
Assreuy
Kyria S
KS
Nascimento
Celso S
CS
Nagano
Benildo S
BS
Cavada
D000595:Amino Acid Sequence; D000818:Animals; D001156:Artemia / Q000187:drug effects / Q000502:physiology; D032449:Canavalia / Q000737:chemistry*; D002846:Chromatography, Affinity; D003911:Dextrans; D004487:Edema / Q000139:chemically induced / Q000276:immunology / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D060748:Fetuins / Q000737:chemistry; D006384:Hemagglutination / Q000187:drug effects; D006614:Hindlimb; D006863:Hydrogen-Ion Concentration; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009005:Monosaccharides / Q000737:chemistry; D010047:Ovalbumin / Q000737:chemistry; D037121:Plant Lectins / Q000302:isolation & purification* / Q000494:pharmacology; D055550:Protein Stability; D021122:Protein Subunits / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012639:Seeds / Q000737:chemistry*
D003911:Dextrans; D060748:Fetuins; D009005:Monosaccharides; D037121:Plant Lectins; D021122:Protein Subunits; C039268:lectin, Canavalia; D010047:Ovalbumin; C025614:sephadex
Canavalia oxyphylla; ESI mass spectrometry; plant lectin; pro-inflammatory; toxic effect