Molecular structure and properties of lectin from tomato fruit.
A cDNA encoding tomato fruit lectin was cloned from an unripe cherry-tomato fruit cDNA library. The isolated lectin cDNA contained an open reading frame encoding 365 amino acids, including peptides that were sequenced. The deduced sequence consisted of three distinct domains: (i) an N-terminal short extensin-like domain; (ii) a Cys-rich carbohydrate binding domain composed of four almost identical chitin-binding domains; (iii) an internal extensin-like domain of 101 residues containing 15 SerPro(4) motifs inserted between the first and second chitin-binding domains. The molecular weight of the lectin was 65,633 and that of the deglycosylated lectin was 32,948, as determined by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). This correlated with the estimated molecular weight of the deduced sequence. Recombinant tomato lectin expressed in Pichia pastoris possessed chitin-binding but not hemagglutinating activity. These findings confirmed that the cDNA encoded tomato lectin.
Oguri
Suguru
S
Department of Bioproduction, Faculty of Bioindustry, Tokyo University of Agriculture, Hokkaido, Japan. s-oguri@bioindustry.nodai.ac.jp
Amano
Koh
K
Nakashita
Hideo
H
Nagata
Yoshiho
Y
Momonoki
Yoshie S
YS
D000595:Amino Acid Sequence; D001483:Base Sequence; D002850:Chromatography, Gel; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D005638:Fruit / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006031:Glycosylation; D018551:Solanum lycopersicum / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D037121:Plant Lectins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018076:DNA, Complementary; D037121:Plant Lectins; D011994:Recombinant Proteins; C034981:tomato lectin