Crystal structures of beta-galactosidase from Penicillium sp. and its complex with galactose

Autor(es)

Rojas, A. L. ^[1] ; Nagem, R. A. P. ^[2] ; Neustroev, K. N. ; Arand, M. ; Adamska, M. ; Eneyskaya, E. V. ; Kulminskaya, A. A. ; Garratt, R. C. ; Golubev, A. M. ; Polikarpov, I. ^[10]

Afiliação do(s) autor(es)

^[1] Universidade de São Paulo (USP). Instituto de Física de São Carlos - Brasil

^[2] Universidade de São Paulo (USP). Instituto de Física de São Carlos - Brasil

^[10] Universidade de São Paulo (USP). Instituto de Física de São Carlos - Brasil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Molecular Biology; v. 343, n. 5, p. 1281-1292, Nov. 2004.
• Área do conhecimento: Ciências Biológicas - Biofísica
• Assunto(s): Biofísica Fungos Enzimas Glicoproteínas Penicillium

Resumo

Beta-galactosidases catalyze the hydrolysis of b(1-3) and b(1-4) galactosyl bonds in oligosaccharides as well as the inverse reaction of enzymatic condensation and transglycosylation. Here we report the crystallographic structures of Penicillium sp. b-galactosidase and its complex with galactose solved by the SIRAS quick cryo-soaking technique at 1.90 A ° and 2.10 A ° resolution, respectively. The amino acid sequence of this 120 kDa protein was first assigned putatively on the basis of inspection of the experimental electron density maps and then determined by nucleotide sequence analysis. Primary structure alignments reveal that Penicillium sp. b-galactosidase belongs to family 35 of glycosyl hydrolases (GHF-35). This model is the first 3D structure for a member of GHF-35. Five distinct domains which comprise the structure are assembled in a way previously unobserved for b-galactosidases. Superposition of this complex with other b-galactosidase complexes from several hydrolase families allowed the identification of residue Glu200 as the proton donor and residue Glu299 as the nucleophile involved in catalysis. Penicillium sp. b-galactosidase is a glycoprotein containing seven N-linked oligosaccharide chains and is the only structure of a glycosylated b-galactosidase described to date. (AU)

• Processo FAPESP: 02/14208-8 - Determinação das estruturas cristalográficas de três glicosidases inéditas e de complexos glicosidases-substrato pelo método de crio derivatização rápida
• Beneficiário: Ronaldo Alves Pinto Nagem
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 01/07014-0 - Estudos estruturais de carbohidrases de fungos
• Beneficiário: Adriana Lucely Rojas Cardona
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 99/03387-4 - Estudos estruturais de proteínas usando luz síncrotron
• Beneficiário: Igor Polikarpov
• Modalidade de apoio: Auxílio à Pesquisa - Temático