Seed lectin from pisum arvense: isolation, biochemical characterization and amino acid sequence.
A glucose/mannose lectin was purified by affinity chromatography from Pisum arvense seeds (PAL) and the 50 kDa molecular mass in solution determined by size exclusion chromatography. SDS-PAGE and electrospray ionization mass spectrometry showed two distinct polypeptide chains: alpha (Mr. 5591 Da) and beta (19986 Da). The lectin was extensively characterized in terms of its biochemical and biological aspects. The amino acid sequence was established by Edman degradation of overlapping peptides. PAL in solution behaves as a dimer and has its monomeric structure formed by two distinct polypeptide chains named alpha (Mr. 5591 Da) and beta (19986 Da) by Electrospray ionization (ESI) mass spectrometry. PAL possesses identical amino acid sequences to that of pea seed lectin but undoubtedly does not exhibit sequence heterogeneity. It is discussed that P. arvense should be considered as a synonym of P. sativum. Furthermore, like pea lectin, PAL discriminates biantennary fucosylated glycan, determined by surface plasmon resonance.
Cavada
Benildo S
BS
Departamento de Bioquímica e Biologia Molecular, Campus do Pici, Caixa Postal 6033, Fortaleza-Ceará. CEP 60451-970, Brazil. bscavada@ufc.br
da Silva
Luzia I M M
LI
Ramos
Márcio V
MV
Galvani
Francisco R
FR
Grangeiro
Thalles B
TB
Leite
Katia B
KB
Assreuy
Ana Maria S
AM
Cajazeiras
Joao B
JB
Calvete
Juan J
JJ
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D019281:Dimerization; D019143:Evolution, Molecular; D007781:Lactoferrin / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018532:Pisum sativum / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D037121:Plant Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D020033:Protein Isoforms / Q000302:isolation & purification / Q000378:metabolism; D012639:Seeds / Q000378:metabolism
D037121:Plant Lectins; D020033:Protein Isoforms; D007781:Lactoferrin