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FEBS J ; 273(17): 3962-74, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16934035

ABSTRACT

Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP-HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407+/-15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N-acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed beta(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-beta-D-glucopyranose units in chitin. The full-length amino acid sequence of Parkia platycephala lectin 2, determined by N-terminal sequencing and cDNA cloning, and its three-dimensional structure, established by X-ray crystallography at 1.75 A resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (betaalpha)8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182.


Subject(s)
Acetylglucosamine/metabolism , Chitinases/chemistry , Fabaceae/enzymology , Hemagglutinins/chemistry , Plant Lectins/chemistry , Seeds/enzymology , Amino Acid Sequence , Base Sequence , Chitinases/genetics , Chitinases/metabolism , Cloning, Molecular , Crystallization , Crystallography, X-Ray , DNA, Complementary/isolation & purification , Fabaceae/genetics , Hemagglutinins/genetics , Hemagglutinins/metabolism , Molecular Sequence Data , Plant Lectins/genetics , Plant Lectins/metabolism , Protein Binding , Seeds/genetics
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