cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N-acetylglucosamine-binding hemagglutinin from Parkia platycephala seeds
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Data
2006-09-01
Autores
Cavada, Benildo S.
Moreno, Frederico B. B. [UNESP]
Da Rocha, Bruno A. M.
De Azevedo Jr., Walter F.
Castellón, Rolando E. R.
Goersch, Georg V.
Nagano, Celso S.
De Souza, Emmanuel P.
Nascimento, Kyria S.
Radis-Baptista, Gandhi
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Resumo
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 β(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-β-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 Å resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (βα) 8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182. © 2006 The Authors.
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Endochitinase, Glycosyl hydrolase family 18, Mimosoideae, Parkia platycephala, X-ray crystal structure, chitin, chitinase, complementary DNA, endochitinase, genomic DNA, glycosidase, hemagglutinin, lectin, lectin 2, n acetylglucosamine, RNA, unclassified drug, affinity chromatography, amino acid composition, amino acid sequence, animal cell, anion exchange chromatography, controlled study, crystal structure, disulfide bond, enzyme activity, enzyme analysis, enzyme inhibition, enzyme purification, erythrocyte, hemagglutination, legume, matrix assisted laser desorption ionization time of flight mass spectrometry, molecular cloning, molecular weight, nonhuman, polyacrylamide gel electrophoresis, priority journal, rabbit, reversed phase high performance liquid chromatography, RNA isolation, sedimentation, X ray crystallography, Acetylglucosamine, Amino Acid Sequence, Base Sequence, Chitinase, Cloning, Molecular, Crystallization, Crystallography, X-Ray, DNA, Complementary, Fabaceae, Hemagglutinins, Molecular Sequence Data, Plant Lectins, Protein Binding, Seeds, Oryctolagus cuniculus
Como citar
FEBS Journal, v. 273, n. 17, p. 3962-3974, 2006.