In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase

Allostery is a central mechanism for the regulation of multi-enzyme complexes. The mechanistic basis that drives allosteric regulation is poorly understood but harbors key information for enzyme engineering. In the present study, we focus on the tryptophan synthase complex that is composed of TrpA a...

Descripción completa

Detalles Bibliográficos
Autores: Maria Solano, Miguel A., Kinateder, Thomas, Iglesias-Fernández, Javier, Sterner, Reinhard, Osuna Oliveras, Sílvia
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10256/20455
Acceso en línea:http://hdl.handle.net/10256/20455
Access Level:acceso abierto
Palabra clave:Triptòfan -- Síntesi
Tryptophan -- Synthesis
Enzims -- Síntesi
Enzymes -- Synthesis
id ES_bc1fc8d4664db582fba11ec742aead13
oai_identifier_str oai:recercat.cat:10256/20455
network_acronym_str ES
network_name_str España
repository_id_str
spelling In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan SynthaseMaria Solano, Miguel A.Kinateder, ThomasIglesias-Fernández, JavierSterner, ReinhardOsuna Oliveras, SílviaTriptòfan -- SíntesiTryptophan -- SynthesisEnzims -- SíntesiEnzymes -- SynthesisAllostery is a central mechanism for the regulation of multi-enzyme complexes. The mechanistic basis that drives allosteric regulation is poorly understood but harbors key information for enzyme engineering. In the present study, we focus on the tryptophan synthase complex that is composed of TrpA and TrpB subunits, which allosterically activate each other. Specifically, we develop a rational approach for identifying key amino acid residues of TrpB distal from the active site. Those residues are predicted to be crucial for shifting the inefficient conformational ensemble of the isolated TrpB to a productive ensemble through intra-subunit allosteric effects. The experimental validation of the conformationally driven TrpB design demonstrates its superior stand-alone activity in the absence of TrpA, comparable to those enhancements obtained after multiple rounds of experimental laboratory evolution. Our work evidences that the current challenge of distal active site prediction for enhanced function in computational enzyme design has become within reachWe thank the Generalitat de Catalunya for the emerging group CompBioLab (2017 SGR-1707) and Spanish MINECO for project PGC2018-102192-B-I00. M.A.M.S. was supported by the Spanish MINECO for a PhD fellowship (BES-2015- 074964) and the National Research Foundation of Korea (NRF) under the Brain Pool Program (NRF2021H1D3A2A02038434), J. I. F. was supported by the European Community for Marie Curie fellowship (H2020- MSCA-IF-2016-753045) and Juan de la Cierva-Incorporación fellowship (IJCI-2017-34129). S.O. is grateful to the funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC-2015-StG-679001) and the Human Frontier Science Program (HFSP) for project grant RGP0054/2020American Chemical Society (ACS)Agencia Estatal de InvestigaciónEuropean Commission2021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionpeer-reviewedapplication/pdfhttp://hdl.handle.net/10256/20455http://hdl.handle.net/10256/20455ACS Catalysis, 2021, vol. 11, núm. 21, p. 13733-13743Articles publicats (D-Q)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)Inglésinfo:eu-repo/semantics/altIdentifier/doi/10.1021/acscatal.1c03950info:eu-repo/semantics/altIdentifier/eissn/2155-5435PGC2018-102192-B-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-102192-B-I00info:eu-repo/grantAgreement/EC/H2020/753045info:eu-repo/grantAgreement/EC/H2020/679001Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10256/204552026-05-29T05:05:01Z
dc.title.none.fl_str_mv In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
title In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
spellingShingle In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
Maria Solano, Miguel A.
Triptòfan -- Síntesi
Tryptophan -- Synthesis
Enzims -- Síntesi
Enzymes -- Synthesis
title_short In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
title_full In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
title_fullStr In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
title_full_unstemmed In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
title_sort In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase
dc.creator.none.fl_str_mv Maria Solano, Miguel A.
Kinateder, Thomas
Iglesias-Fernández, Javier
Sterner, Reinhard
Osuna Oliveras, Sílvia
author Maria Solano, Miguel A.
author_facet Maria Solano, Miguel A.
Kinateder, Thomas
Iglesias-Fernández, Javier
Sterner, Reinhard
Osuna Oliveras, Sílvia
author_role author
author2 Kinateder, Thomas
Iglesias-Fernández, Javier
Sterner, Reinhard
Osuna Oliveras, Sílvia
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Agencia Estatal de Investigación
European Commission
dc.subject.none.fl_str_mv Triptòfan -- Síntesi
Tryptophan -- Synthesis
Enzims -- Síntesi
Enzymes -- Synthesis
topic Triptòfan -- Síntesi
Tryptophan -- Synthesis
Enzims -- Síntesi
Enzymes -- Synthesis
description Allostery is a central mechanism for the regulation of multi-enzyme complexes. The mechanistic basis that drives allosteric regulation is poorly understood but harbors key information for enzyme engineering. In the present study, we focus on the tryptophan synthase complex that is composed of TrpA and TrpB subunits, which allosterically activate each other. Specifically, we develop a rational approach for identifying key amino acid residues of TrpB distal from the active site. Those residues are predicted to be crucial for shifting the inefficient conformational ensemble of the isolated TrpB to a productive ensemble through intra-subunit allosteric effects. The experimental validation of the conformationally driven TrpB design demonstrates its superior stand-alone activity in the absence of TrpA, comparable to those enhancements obtained after multiple rounds of experimental laboratory evolution. Our work evidences that the current challenge of distal active site prediction for enhanced function in computational enzyme design has become within reach
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
peer-reviewed
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10256/20455
http://hdl.handle.net/10256/20455
url http://hdl.handle.net/10256/20455
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1021/acscatal.1c03950
info:eu-repo/semantics/altIdentifier/eissn/2155-5435
PGC2018-102192-B-I00
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-102192-B-I00
info:eu-repo/grantAgreement/EC/H2020/753045
info:eu-repo/grantAgreement/EC/H2020/679001
dc.rights.none.fl_str_mv Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Chemical Society (ACS)
publisher.none.fl_str_mv American Chemical Society (ACS)
dc.source.none.fl_str_mv ACS Catalysis, 2021, vol. 11, núm. 21, p. 13733-13743
Articles publicats (D-Q)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869418085981618176
score 15.812429

Similares en LA Referencia