Crystal structure of α-galactosidase from Trichoderma reesei and its complex with galactose: Implications for catalytic mechanism. Golubev, A., Nagem, R., Brandão Neto, J., Neustroev, K., Eneyskaya, E., Kulminskaya, A., Shabalin, K., Savel'ev, A., & Polikarpov, I. Journal of Molecular Biology, 339(2):413-422, 2004. cited By 60![Crystal structure of α-galactosidase from Trichoderma reesei and its complex with galactose: Implications for catalytic mechanism [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex The crystal structures of α-galactosidase from the mesophilic fungus Trichoderma reesei and its complex with the competitive inhibitor, β-D-galactose, have been determined at 1.54Å and 2.0Å resolution, respectively. The α-galactosidase structure was solved by the quick cryo-soaking method using a single Cs derivative. The refined crystallographic model of the α-galactosidase consists of two domains, an N-terminal catalytic domain of the (β/α)8 barrel topology and a C-terminal domain which is formed by an antiparallel β-structure. The protein contains four N-glycosylation sites located in the catalytic domain. Some of the oligosaccharides were found to participate in inter-domain contacts. The galactose molecule binds to the active site pocket located in the center of the barrel of the catalytic domain. Analysis of the α-galactosidase- galactose complex reveals the residues of the active site and offers a structural basis for identification of the putative mechanism of the enzymatic reaction. The structure of the α-galactosidase closely resembles those of the glycoside hydrolase family 27. The conservation of two catalytic Asp residues, identified for this family, is consistent with a double-displacement reaction mechanism for the α-galactosidase. Modeling of possible substrates into the active site reveals specific hydrogen bonds and hydrophobic interactions that could explain peculiarities of the enzyme kinetics. © 2004 Elsevier Ltd. All rights reserved.
@ARTICLE{Golubev2004413,
author={Golubev, A.M. and Nagem, R.A.P. and Brandão Neto, J.R. and Neustroev, K.N. and Eneyskaya, E.V. and Kulminskaya, A.A. and Shabalin, K.A. and Savel'ev, A.N. and Polikarpov, I.},
title={Crystal structure of α-galactosidase from Trichoderma reesei and its complex with galactose: Implications for catalytic mechanism},
journal={Journal of Molecular Biology},
year={2004},
volume={339},
number={2},
pages={413-422},
doi={10.1016/j.jmb.2004.03.062},
note={cited By 60},
url={https://www.scopus.com/inward/record.uri?eid=2-s2.0-2342515384&doi=10.1016%2fj.jmb.2004.03.062&partnerID=40&md5=7b74bf3ff3c0db547bf566e37d2a41b9},
affiliation={Petersburg Nuclear Physics Institute, Gatchina, St Petersburg, 188300, Russian Federation; Instituto de Física, Universidade de São Paulo, Av. Trabalhador Saocarlense, 400, CEP 13560-970, São Carlos, SP, Brazil; Biophysics Department, St. Petersburg Technical University, 29 Politechnicheskaya St., St Petersburg, 195251, Russian Federation},
abstract={The crystal structures of α-galactosidase from the mesophilic fungus Trichoderma reesei and its complex with the competitive inhibitor, β-D-galactose, have been determined at 1.54Å and 2.0Å resolution, respectively. The α-galactosidase structure was solved by the quick cryo-soaking method using a single Cs derivative. The refined crystallographic model of the α-galactosidase consists of two domains, an N-terminal catalytic domain of the (β/α)8 barrel topology and a C-terminal domain which is formed by an antiparallel β-structure. The protein contains four N-glycosylation sites located in the catalytic domain. Some of the oligosaccharides were found to participate in inter-domain contacts. The galactose molecule binds to the active site pocket located in the center of the barrel of the catalytic domain. Analysis of the α-galactosidase- galactose complex reveals the residues of the active site and offers a structural basis for identification of the putative mechanism of the enzymatic reaction. The structure of the α-galactosidase closely resembles those of the glycoside hydrolase family 27. The conservation of two catalytic Asp residues, identified for this family, is consistent with a double-displacement reaction mechanism for the α-galactosidase. Modeling of possible substrates into the active site reveals specific hydrogen bonds and hydrophobic interactions that could explain peculiarities of the enzyme kinetics. © 2004 Elsevier Ltd. All rights reserved.},
author_keywords={α-GAL, α-galactosidase; α-galactosidase; galactose; GHF, glycoside hydrolase family; N-glycosylation; oxidative activation; SIRAS, single isomorphous replacement with anomalous scattering; Trichoderma reesei},
funding_details={Fundação de Amparo à Pesquisa do Estado de São Paulo03-04-48756},
funding_details={99/03387-4, 02/14208-8},
}
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All rights reserved.","author_keywords":"α-GAL, α-galactosidase; α-galactosidase; galactose; GHF, glycoside hydrolase family; N-glycosylation; oxidative activation; SIRAS, single isomorphous replacement with anomalous scattering; Trichoderma reesei","funding_details":"99/03387-4, 02/14208-8","bibtex":"@ARTICLE{Golubev2004413,\r\nauthor={Golubev, A.M. and Nagem, R.A.P. and Brandão Neto, J.R. and Neustroev, K.N. and Eneyskaya, E.V. and Kulminskaya, A.A. and Shabalin, K.A. and Savel'ev, A.N. and Polikarpov, I.},\r\ntitle={Crystal structure of α-galactosidase from Trichoderma reesei and its complex with galactose: Implications for catalytic mechanism},\r\njournal={Journal of Molecular Biology},\r\nyear={2004},\r\nvolume={339},\r\nnumber={2},\r\npages={413-422},\r\ndoi={10.1016/j.jmb.2004.03.062},\r\nnote={cited By 60},\r\nurl={https://www.scopus.com/inward/record.uri?eid=2-s2.0-2342515384&doi=10.1016%2fj.jmb.2004.03.062&partnerID=40&md5=7b74bf3ff3c0db547bf566e37d2a41b9},\r\naffiliation={Petersburg Nuclear Physics Institute, Gatchina, St Petersburg, 188300, Russian Federation; Instituto de Física, Universidade de São Paulo, Av. 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