Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.procbio.2022.12.010 http://hdl.handle.net/11449/246511 |
Resumo: | Leptospirosis is a bacterial disease that affects humans and animals and is caused by Leptospira. The recommended treatment for leptospirosis is antibiotic therapy, which should be given early in the course of the disease. Despite the use of these antibiotics, their role during the course of the disease is still not completely clear because of the lack of effective clinical trials, particularly for severe cases of the disease. Here, we present the characterization of L. interrogans Lsa45 protein by gel filtration, protein crystallography, SAXS, fluorescence and enzymatic assays. The oligomeric studies revealed that Lsa45 is monomeric in solution. The crystal structure of Lsa45 revealed the presence of two subdomains: a large α/β subdomain and a small α-helical subdomain. The large subdomain contains the amino acids Ser122, Lys125, and Tyr217, which correspond to the catalytic triad that is essential for β-lactamase or serine hydrolase activity in similar enzymes. Additionally, we also confirmed the bifunctional promiscuity of Lsa45, in hydrolyzing both the 4-nitrophenyl acetate (p-NPA) and nitrocefin β-lactam antibiotic. Therefore, this study provides novel insights into the structure and function of enzymes from L. interrogans, which furthers our understanding of this bacterium and the development of new therapies for the prevention and treatment of leptospirosis. |
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Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activityBifunctional enzymeCrystal structureL. interrogansLeptospirosisPBPβ-lactamase/esteraseLeptospirosis is a bacterial disease that affects humans and animals and is caused by Leptospira. The recommended treatment for leptospirosis is antibiotic therapy, which should be given early in the course of the disease. Despite the use of these antibiotics, their role during the course of the disease is still not completely clear because of the lack of effective clinical trials, particularly for severe cases of the disease. Here, we present the characterization of L. interrogans Lsa45 protein by gel filtration, protein crystallography, SAXS, fluorescence and enzymatic assays. The oligomeric studies revealed that Lsa45 is monomeric in solution. The crystal structure of Lsa45 revealed the presence of two subdomains: a large α/β subdomain and a small α-helical subdomain. The large subdomain contains the amino acids Ser122, Lys125, and Tyr217, which correspond to the catalytic triad that is essential for β-lactamase or serine hydrolase activity in similar enzymes. Additionally, we also confirmed the bifunctional promiscuity of Lsa45, in hydrolyzing both the 4-nitrophenyl acetate (p-NPA) and nitrocefin β-lactam antibiotic. Therefore, this study provides novel insights into the structure and function of enzymes from L. interrogans, which furthers our understanding of this bacterium and the development of new therapies for the prevention and treatment of leptospirosis.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)National Institutes of HealthLaboratório de Desenvolvimento de Vacinas Instituto Butantan, Avenida Vital Brasil, 1500, SPDepartment of Chemistry & Biochemistry University of CaliforniaDepartamento de Bioquímica e Imunologia Instituto de Ciências Biológicas (ICB) Universidade Federal de Minas Gerais (UFMG)Universidade Estadual Paulista (UNESP) Instituto de Biociências Dep. de Física e Biofísica, SPInstituto Federal da Bahia – IFBA, Rodovia BR-367, R. José Fontana, 1, BAUniversidade Estadual Paulista (UNESP) Instituto de Biociências Dep. de Física e Biofísica, SPFAPESP: 2014/50981-0FAPESP: 2017/06731-8FAPESP: 2019/17488-2CNPq: 301229/2017-1CNPq: 304445/2021-5National Institutes of Health: R56 AI096837Instituto ButantanUniversity of CaliforniaUniversidade Federal de Minas Gerais (UFMG)Universidade Estadual Paulista (UNESP)Instituto Federal da Bahia – IFBASantos, Jademilson C.Handa, SumitFernandes, Luis G.V.Bleicher, LucasGandin, César A. [UNESP]de Oliveira-Neto, Mario [UNESP]Ghosh, ParthoNascimento, Ana Lucia T.O.2023-07-29T12:42:59Z2023-07-29T12:42:59Z2023-02-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article141-153http://dx.doi.org/10.1016/j.procbio.2022.12.010Process Biochemistry, v. 125, p. 141-153.1359-5113http://hdl.handle.net/11449/24651110.1016/j.procbio.2022.12.0102-s2.0-85144505086Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengProcess Biochemistryinfo:eu-repo/semantics/openAccess2023-07-29T12:42:59Zoai:repositorio.unesp.br:11449/246511Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:16:36.034451Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
title |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
spellingShingle |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity Santos, Jademilson C. Bifunctional enzyme Crystal structure L. interrogans Leptospirosis PBP β-lactamase/esterase |
title_short |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
title_full |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
title_fullStr |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
title_full_unstemmed |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
title_sort |
Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity |
author |
Santos, Jademilson C. |
author_facet |
Santos, Jademilson C. Handa, Sumit Fernandes, Luis G.V. Bleicher, Lucas Gandin, César A. [UNESP] de Oliveira-Neto, Mario [UNESP] Ghosh, Partho Nascimento, Ana Lucia T.O. |
author_role |
author |
author2 |
Handa, Sumit Fernandes, Luis G.V. Bleicher, Lucas Gandin, César A. [UNESP] de Oliveira-Neto, Mario [UNESP] Ghosh, Partho Nascimento, Ana Lucia T.O. |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Instituto Butantan University of California Universidade Federal de Minas Gerais (UFMG) Universidade Estadual Paulista (UNESP) Instituto Federal da Bahia – IFBA |
dc.contributor.author.fl_str_mv |
Santos, Jademilson C. Handa, Sumit Fernandes, Luis G.V. Bleicher, Lucas Gandin, César A. [UNESP] de Oliveira-Neto, Mario [UNESP] Ghosh, Partho Nascimento, Ana Lucia T.O. |
dc.subject.por.fl_str_mv |
Bifunctional enzyme Crystal structure L. interrogans Leptospirosis PBP β-lactamase/esterase |
topic |
Bifunctional enzyme Crystal structure L. interrogans Leptospirosis PBP β-lactamase/esterase |
description |
Leptospirosis is a bacterial disease that affects humans and animals and is caused by Leptospira. The recommended treatment for leptospirosis is antibiotic therapy, which should be given early in the course of the disease. Despite the use of these antibiotics, their role during the course of the disease is still not completely clear because of the lack of effective clinical trials, particularly for severe cases of the disease. Here, we present the characterization of L. interrogans Lsa45 protein by gel filtration, protein crystallography, SAXS, fluorescence and enzymatic assays. The oligomeric studies revealed that Lsa45 is monomeric in solution. The crystal structure of Lsa45 revealed the presence of two subdomains: a large α/β subdomain and a small α-helical subdomain. The large subdomain contains the amino acids Ser122, Lys125, and Tyr217, which correspond to the catalytic triad that is essential for β-lactamase or serine hydrolase activity in similar enzymes. Additionally, we also confirmed the bifunctional promiscuity of Lsa45, in hydrolyzing both the 4-nitrophenyl acetate (p-NPA) and nitrocefin β-lactam antibiotic. Therefore, this study provides novel insights into the structure and function of enzymes from L. interrogans, which furthers our understanding of this bacterium and the development of new therapies for the prevention and treatment of leptospirosis. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-07-29T12:42:59Z 2023-07-29T12:42:59Z 2023-02-01 |
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 |
http://dx.doi.org/10.1016/j.procbio.2022.12.010 Process Biochemistry, v. 125, p. 141-153. 1359-5113 http://hdl.handle.net/11449/246511 10.1016/j.procbio.2022.12.010 2-s2.0-85144505086 |
url |
http://dx.doi.org/10.1016/j.procbio.2022.12.010 http://hdl.handle.net/11449/246511 |
identifier_str_mv |
Process Biochemistry, v. 125, p. 141-153. 1359-5113 10.1016/j.procbio.2022.12.010 2-s2.0-85144505086 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Process Biochemistry |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
141-153 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808128782133886976 |