Mostrar el registro sencillo del ítem

dc.contributor.author
Paris, Gastón
dc.contributor.author
Ratier, Laura
dc.contributor.author
Amaya, María Fernanda
dc.contributor.author
Nguyen, Tong
dc.contributor.author
Alzari, Pedro M.
dc.contributor.author
Frasch, Alberto Carlos C.
dc.date.available
2018-03-21T18:54:32Z
dc.date.issued
2005-01
dc.identifier.citation
Paris, Gastón; Ratier, Laura; Amaya, María Fernanda; Nguyen, Tong; Alzari, Pedro M.; et al.; A sialidase mutant displaying trans-sialidase activity; Elsevier; Journal Of Molecular Biology; 345; 4; 1-2005; 923-934
dc.identifier.issn
0022-2836
dc.identifier.uri
http://hdl.handle.net/11336/39546
dc.description.abstract
Trypanosoma cruzi, the agent of Chagas disease, expresses a modified sialidase, the trans-sialidase, which transfers sialic acid from host glycoconjugates to β-galactose present in parasite mucins. Another American trypanosome, Trypanosoma rangeli, expresses a homologous protein that has sialidase activity but is devoid of transglycosidase activity. Based on the recently determined structures of T. rangeli sialidase (TrSA) and T. cruzi trans-sialidase (TcTS), we have now constructed mutants of TrSA with the aim of studing the relevant residues in transfer activity. Five mutations, Met96-Val, Ala98-Pro, Ser120-Tyr, Gly249-Tyr and Gln284-Pro, were enough to obtain a sialidase mutant (TrSA 5mut) with trans-sialidase activity; and a sixth mutation increased the activity to about 10% that of wild-type TcTS. The crystal structure of TrSA 5mut revealed the formation of a trans-sialidase-like binding site for the acceptor galactose, primarily defined by the phenol group of Tyr120 and the indole ring of Trp313, which adopts a new conformation, similar to that in TcTS, induced by the Gln284-Pro mutation. The transition state analogue 2,3-didehydro-2-deoxy-N-acetylneuraminic acid (DANA), which inhibits sialidases but is a poor inhibitor of trans-sialidase, was used to probe the active site conformation of mutant enzymes. The results show that the presence of a sugar acceptor binding-site, the fine-tuning of protein-substrate interactions and the flexibility of crucial active site residues are all important to achieve transglycosidase activity from the TrSA sialidase scaffold.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Protein Engineering
dc.subject
Sialidase
dc.subject
Trans-Sialidase
dc.subject
Trypanosoma Cruzi
dc.subject
Trypanosoma Rangeli
dc.subject.classification
Otras Ciencias Biológicas
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
A sialidase mutant displaying trans-sialidase activity
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-03-15T18:03:36Z
dc.identifier.eissn
1089-8638
dc.journal.volume
345
dc.journal.number
4
dc.journal.pagination
923-934
dc.journal.pais
Países Bajos
dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Paris, Gastón. Universidad Nacional de San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
dc.description.fil
Fil: Ratier, Laura. Universidad Nacional de San Martín; Argentina
dc.description.fil
Fil: Amaya, María Fernanda. Instituto Pasteur; Francia
dc.description.fil
Fil: Nguyen, Tong. Instituto Pasteur; Francia
dc.description.fil
Fil: Alzari, Pedro M.. Instituto Pasteur; Francia
dc.description.fil
Fil: Frasch, Alberto Carlos C.. Universidad Nacional de San Martín; Argentina
dc.journal.title
Journal Of Molecular Biology
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S002228360401174X
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.jmb.2004.09.031


Archivos asociados

Documento no disponible

Comunidades y colecciones

Mostrar el registro sencillo del ítem