Artigo
cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N-acetylglucosamine-binding hemagglutinin from Parkia platycephala seeds
Fecha
2006-09-01Registro en:
FEBS Journal, v. 273, n. 17, p. 3962-3974, 2006.
1742-464X
1742-4658
10.1111/j.1742-4658.2006.05400.x
2-s2.0-33747413686
2-s2.0-33747413686.pdf
Autor
Universidade Federal do Ceará (UFC)
Universidade Estadual Paulista (Unesp)
Universidade Regional Do Cariri
Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)
CSIC
Université des Sciences et Technologies de Lille
Universidade Estadual de Campinas (UNICAMP)
Bâtiment C-9
Resumen
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.