Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/10216/136227 |
Resumo: | Laccases (EC 1.10.3.2) are multicopper oxidoreductases acting on diphenols and related substances. Laccases are highly important for biotechnology and environmental remediation. These enzymes contain mononuclear one T2 copper ion and two T3 copper ions (Cu3a and Cu3ß), which form the so-called trinuclear center (TNC). Along with the typical three-domain laccases Bacteria produce two-domain (2D) enzymes, which are active at neutral and basic pH, thermostable, and resistant to inhibitors. In this work we present the comparative analysis of crystal structures and catalytic properties of recombinant 2D laccase from Streptomyces griseoflavus Ac-993 (SgfSL) and its four mutant forms with replacements of two amino acid residues, located at the narrowing of the presumable T3-solvent tunnels. We obtained inactive enzymes with substitutions of His165, with Phe, and Ile170 with Ala or Phe. His165Ala variant was more active than the wild type. We suggest that His 165 is a “gateway” at the O2-tunnel leading from solvent to the Cu3ß of the enzyme. The side chain of Ile170 could be indirectly involved in the coordination of copper ions at the T3 center by maintaining the position of the imidazole ring of His 157 that belongs to the first coordination sphere of Cu3a. |
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Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993Crystal structuresT2/T3 copper siteTunnelsTwo-domain laccasesLaccases (EC 1.10.3.2) are multicopper oxidoreductases acting on diphenols and related substances. Laccases are highly important for biotechnology and environmental remediation. These enzymes contain mononuclear one T2 copper ion and two T3 copper ions (Cu3a and Cu3ß), which form the so-called trinuclear center (TNC). Along with the typical three-domain laccases Bacteria produce two-domain (2D) enzymes, which are active at neutral and basic pH, thermostable, and resistant to inhibitors. In this work we present the comparative analysis of crystal structures and catalytic properties of recombinant 2D laccase from Streptomyces griseoflavus Ac-993 (SgfSL) and its four mutant forms with replacements of two amino acid residues, located at the narrowing of the presumable T3-solvent tunnels. We obtained inactive enzymes with substitutions of His165, with Phe, and Ile170 with Ala or Phe. His165Ala variant was more active than the wild type. We suggest that His 165 is a “gateway” at the O2-tunnel leading from solvent to the Cu3ß of the enzyme. The side chain of Ile170 could be indirectly involved in the coordination of copper ions at the T3 center by maintaining the position of the imidazole ring of His 157 that belongs to the first coordination sphere of Cu3a.MDPI20192019-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10216/136227eng1661-659610.3390/ijms20133184Gabdulkhakov, AKolyadenko, IKostareva, OMikhaylina, AOliveira, PTamagnini, PLisov, ATishchenko, Sinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-29T14:58:46Zoai:repositorio-aberto.up.pt:10216/136227Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:12:53.075704Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
title |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
spellingShingle |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 Gabdulkhakov, A Crystal structures T2/T3 copper site Tunnels Two-domain laccases |
title_short |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
title_full |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
title_fullStr |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
title_full_unstemmed |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
title_sort |
Investigations of accessibility of T2/T3 copper center of two-domain laccase from streptomyces griseoflavus ac-993 |
author |
Gabdulkhakov, A |
author_facet |
Gabdulkhakov, A Kolyadenko, I Kostareva, O Mikhaylina, A Oliveira, P Tamagnini, P Lisov, A Tishchenko, S |
author_role |
author |
author2 |
Kolyadenko, I Kostareva, O Mikhaylina, A Oliveira, P Tamagnini, P Lisov, A Tishchenko, S |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Gabdulkhakov, A Kolyadenko, I Kostareva, O Mikhaylina, A Oliveira, P Tamagnini, P Lisov, A Tishchenko, S |
dc.subject.por.fl_str_mv |
Crystal structures T2/T3 copper site Tunnels Two-domain laccases |
topic |
Crystal structures T2/T3 copper site Tunnels Two-domain laccases |
description |
Laccases (EC 1.10.3.2) are multicopper oxidoreductases acting on diphenols and related substances. Laccases are highly important for biotechnology and environmental remediation. These enzymes contain mononuclear one T2 copper ion and two T3 copper ions (Cu3a and Cu3ß), which form the so-called trinuclear center (TNC). Along with the typical three-domain laccases Bacteria produce two-domain (2D) enzymes, which are active at neutral and basic pH, thermostable, and resistant to inhibitors. In this work we present the comparative analysis of crystal structures and catalytic properties of recombinant 2D laccase from Streptomyces griseoflavus Ac-993 (SgfSL) and its four mutant forms with replacements of two amino acid residues, located at the narrowing of the presumable T3-solvent tunnels. We obtained inactive enzymes with substitutions of His165, with Phe, and Ile170 with Ala or Phe. His165Ala variant was more active than the wild type. We suggest that His 165 is a “gateway” at the O2-tunnel leading from solvent to the Cu3ß of the enzyme. The side chain of Ile170 could be indirectly involved in the coordination of copper ions at the T3 center by maintaining the position of the imidazole ring of His 157 that belongs to the first coordination sphere of Cu3a. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019 2019-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://hdl.handle.net/10216/136227 |
url |
https://hdl.handle.net/10216/136227 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1661-6596 10.3390/ijms20133184 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
MDPI |
publisher.none.fl_str_mv |
MDPI |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799136051006013440 |