β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico
Ano de defesa: | 2007 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Santa Maria
|
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
|
Departamento: |
Bioquímica
|
País: |
BR
|
Palavras-chave em Português: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://repositorio.ufsm.br/handle/1/11085 |
Resumo: | Methylmalonic acidemia, one of the most frequent organic acidemias, is caused by deficiency of the methylmalonyl CoA mutase, leading to tissue accumulation of Lmethylmalolonic acid (MMA). Affects individuals present lethargy, coma, vomiting, muscular hypotonia, recurrent episodes of metabolic acidosis and progressive encephalopathy. In this context, it has been proposed that MMA inhibits succinate dehydrogenase (SDH) and leads to ATP depletion, lactate accumulation and excitotoxic damage. In the present study we confirmed that MMA competitively inhibits of SDH, and later were investigated the enzymatic medium reduction alters the activity of SDH or its inhibition by MMA. SDH activity was determined in cerebral cortex homogenates, using 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT), as the electorn acceptor. The reduction enzymatic medium was promoted by preincubation with β-NADH (160 μM). The preincubation of β-NADH prevented the inhibition of the activity of SDH induzed by 5mM of the MMA [F(1,5)=9.31; p=0.028]. In addition, was determined of the kinetic parameters (Km and Vmax) and inhition constant (Ki ) of SDH preincubated in the presence and absence of β-NADH. The Km of SDH preincubated with β-NADH (Km=0.216 nmol INT) was different of the Km of SDH preincubated in the absence of the β-NADH (Km=0.272nmol INT) [T(2)=10.375; p=0.009]. The presence of β-NADH in the incubation medium did not alter Vmax= 4.72 ± 0.28.10-8 mol INT/mg protein/min) [T(2)=-1.0; p=0.423]. The Ki of the MMA by SDH activity in presence of the β- NADH (Ki =20.05 mM) is increased that of Ki of the MMA by SDH activity in absence of the β-NADH (Ki =11.60 mM), [T(2)=18.806; p=0.003]. In conclusion, we showed that, in the presence of β-NADH, SDH is less sensitive to the inhibition by MMA. |
id |
UFSM-20_b4429a4590fffa1a06f839bea67215ca |
---|---|
oai_identifier_str |
oai:repositorio.ufsm.br:1/11085 |
network_acronym_str |
UFSM-20 |
network_name_str |
Manancial - Repositório Digital da UFSM |
repository_id_str |
|
spelling |
2017-04-112017-04-112007-05-04TORRES, Aledson Rosa. β-Nadh reduces succinate dehydrogenase sensitivity to the competitive inhibitor methylmalonic acid in cerebral cortex of rats. 2007. 72 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2007.http://repositorio.ufsm.br/handle/1/11085Methylmalonic acidemia, one of the most frequent organic acidemias, is caused by deficiency of the methylmalonyl CoA mutase, leading to tissue accumulation of Lmethylmalolonic acid (MMA). Affects individuals present lethargy, coma, vomiting, muscular hypotonia, recurrent episodes of metabolic acidosis and progressive encephalopathy. In this context, it has been proposed that MMA inhibits succinate dehydrogenase (SDH) and leads to ATP depletion, lactate accumulation and excitotoxic damage. In the present study we confirmed that MMA competitively inhibits of SDH, and later were investigated the enzymatic medium reduction alters the activity of SDH or its inhibition by MMA. SDH activity was determined in cerebral cortex homogenates, using 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT), as the electorn acceptor. The reduction enzymatic medium was promoted by preincubation with β-NADH (160 μM). The preincubation of β-NADH prevented the inhibition of the activity of SDH induzed by 5mM of the MMA [F(1,5)=9.31; p=0.028]. In addition, was determined of the kinetic parameters (Km and Vmax) and inhition constant (Ki ) of SDH preincubated in the presence and absence of β-NADH. The Km of SDH preincubated with β-NADH (Km=0.216 nmol INT) was different of the Km of SDH preincubated in the absence of the β-NADH (Km=0.272nmol INT) [T(2)=10.375; p=0.009]. The presence of β-NADH in the incubation medium did not alter Vmax= 4.72 ± 0.28.10-8 mol INT/mg protein/min) [T(2)=-1.0; p=0.423]. The Ki of the MMA by SDH activity in presence of the β- NADH (Ki =20.05 mM) is increased that of Ki of the MMA by SDH activity in absence of the β-NADH (Ki =11.60 mM), [T(2)=18.806; p=0.003]. In conclusion, we showed that, in the presence of β-NADH, SDH is less sensitive to the inhibition by MMA.A acidemia metilmalônica, uma das mais frequentes acidemias orgânicas, é causada pela deficiência da enzima metilmalonil-CoA mutase, ocasionando acumulação tecidual de ácido L-metilmalônico (MMA). Os indivíduos afetados apresentam letargia, coma, vômito, hipotonia muscular, episódios recorrentes de acidose metabólica e encefalopatia progressiva. Em decorrência do acúmulo de MMA ocorre inibição da sucinato desidrogenase (SDH), E.C.5.5.99.2, diminuição da produção de ATP, aumento dos níveis de lactato e dano excitotóxico. No presente estudo confirmamos que o MMA inibe a SDH competitivamente e investigamos se a presença de uma agente redutor no meio enzimático alteraria a atividade da SDH, ou sua inibição por MMA. A atividade da SDH em córtex cerebral de ratos foi determinada usando o 2-(p-iodofenil)-3-(p-nitrofenil)-5-feniltetrazólio (INT), como aceptor de elétrons. A pré-incubação com β-NADH (160 μM) preveniu a inibição da SDH causada por 5 mM de MMA [F(1,5)=9,31; p=0,028]. Posteriormente, foi determinado os parâmetros cinéticos (Km e Vmax) da SDH pré-incubada na presença e ausência de β- NADH. O Km da SDH pré-incubada com β-NADH (Km=0,216 nmol INT) é menor que o Km da SDH pré-incubada na ausência de β-NADH (Km=0,272nmol INT), [T(2)=10,375; p=0,009]. Por outro lado a pré-incubação da enzima com β-NADH não alterou a Vmax (4,72 ± 0,28.10-8 mol INT/mg proteína/min) [T(2)=-1,0; p=0,423]. A constante de inibição (Ki) do MMA para a SDH pré-incubada com β-NADH (Ki=20,05 mM) foi maior que o Ki do MMA para a SDH préincubada sem β-NADH (Ki=11,60 mM) [T(2)=18,806; p=0,003]. Os dados mostram que a presença de β-NADH diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico.application/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaUFSMBRBioquímicaBioquímicaEnzimasÁcido metilmalônicoSucinato desidrogenaseCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAβ-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônicoβ-Nadh reduces succinate dehydrogenase sensitivity to the competitive inhibitor methylmalonic acid in cerebral cortex of ratsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisMello, Carlos Fernando dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782674D2Coitinho, Adriana Simonhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4768906A0Pereira, Maria Esterhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4728086Y2http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4103709P6Torres, Aledson Rosa20080000000240050030030050082e537b0-4a43-400e-9f82-0cd91952adeaf883e09b-a927-4fe9-b1a2-7b29bde49b194558062d-b26b-4b7d-9adf-5ba876b07d08276bcfc2-822c-4d05-88f0-be6c067a20c1info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALALEDSONTORRES.pdfapplication/pdf2173344http://repositorio.ufsm.br/bitstream/1/11085/1/ALEDSONTORRES.pdff964591d9a022846aed4a3182d8f88a7MD51TEXTALEDSONTORRES.pdf.txtALEDSONTORRES.pdf.txtExtracted texttext/plain84995http://repositorio.ufsm.br/bitstream/1/11085/2/ALEDSONTORRES.pdf.txta6ff7b8900fd7cf8876e829bc9f4852dMD52THUMBNAILALEDSONTORRES.pdf.jpgALEDSONTORRES.pdf.jpgIM Thumbnailimage/jpeg4759http://repositorio.ufsm.br/bitstream/1/11085/3/ALEDSONTORRES.pdf.jpgce39cf9d37610fa334816b00d8529430MD531/110852017-07-25 12:09:59.74oai:repositorio.ufsm.br:1/11085Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132017-07-25T15:09:59Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
dc.title.alternative.eng.fl_str_mv |
β-Nadh reduces succinate dehydrogenase sensitivity to the competitive inhibitor methylmalonic acid in cerebral cortex of rats |
title |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
spellingShingle |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico Torres, Aledson Rosa Bioquímica Enzimas Ácido metilmalônico Sucinato desidrogenase CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
title_short |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
title_full |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
title_fullStr |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
title_full_unstemmed |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
title_sort |
β-nadh diminui a sensibilidade da sucinato desidrogenase a inibição por ácido metilmalônico |
author |
Torres, Aledson Rosa |
author_facet |
Torres, Aledson Rosa |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Mello, Carlos Fernando de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782674D2 |
dc.contributor.referee1.fl_str_mv |
Coitinho, Adriana Simon |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4768906A0 |
dc.contributor.referee2.fl_str_mv |
Pereira, Maria Ester |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4728086Y2 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4103709P6 |
dc.contributor.author.fl_str_mv |
Torres, Aledson Rosa |
contributor_str_mv |
Mello, Carlos Fernando de Coitinho, Adriana Simon Pereira, Maria Ester |
dc.subject.por.fl_str_mv |
Bioquímica Enzimas Ácido metilmalônico Sucinato desidrogenase |
topic |
Bioquímica Enzimas Ácido metilmalônico Sucinato desidrogenase CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
description |
Methylmalonic acidemia, one of the most frequent organic acidemias, is caused by deficiency of the methylmalonyl CoA mutase, leading to tissue accumulation of Lmethylmalolonic acid (MMA). Affects individuals present lethargy, coma, vomiting, muscular hypotonia, recurrent episodes of metabolic acidosis and progressive encephalopathy. In this context, it has been proposed that MMA inhibits succinate dehydrogenase (SDH) and leads to ATP depletion, lactate accumulation and excitotoxic damage. In the present study we confirmed that MMA competitively inhibits of SDH, and later were investigated the enzymatic medium reduction alters the activity of SDH or its inhibition by MMA. SDH activity was determined in cerebral cortex homogenates, using 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT), as the electorn acceptor. The reduction enzymatic medium was promoted by preincubation with β-NADH (160 μM). The preincubation of β-NADH prevented the inhibition of the activity of SDH induzed by 5mM of the MMA [F(1,5)=9.31; p=0.028]. In addition, was determined of the kinetic parameters (Km and Vmax) and inhition constant (Ki ) of SDH preincubated in the presence and absence of β-NADH. The Km of SDH preincubated with β-NADH (Km=0.216 nmol INT) was different of the Km of SDH preincubated in the absence of the β-NADH (Km=0.272nmol INT) [T(2)=10.375; p=0.009]. The presence of β-NADH in the incubation medium did not alter Vmax= 4.72 ± 0.28.10-8 mol INT/mg protein/min) [T(2)=-1.0; p=0.423]. The Ki of the MMA by SDH activity in presence of the β- NADH (Ki =20.05 mM) is increased that of Ki of the MMA by SDH activity in absence of the β-NADH (Ki =11.60 mM), [T(2)=18.806; p=0.003]. In conclusion, we showed that, in the presence of β-NADH, SDH is less sensitive to the inhibition by MMA. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007-05-04 |
dc.date.accessioned.fl_str_mv |
2017-04-11 |
dc.date.available.fl_str_mv |
2017-04-11 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
TORRES, Aledson Rosa. β-Nadh reduces succinate dehydrogenase sensitivity to the competitive inhibitor methylmalonic acid in cerebral cortex of rats. 2007. 72 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2007. |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/11085 |
identifier_str_mv |
TORRES, Aledson Rosa. β-Nadh reduces succinate dehydrogenase sensitivity to the competitive inhibitor methylmalonic acid in cerebral cortex of rats. 2007. 72 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2007. |
url |
http://repositorio.ufsm.br/handle/1/11085 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
200800000002 |
dc.relation.confidence.fl_str_mv |
400 500 300 300 500 |
dc.relation.authority.fl_str_mv |
82e537b0-4a43-400e-9f82-0cd91952adea f883e09b-a927-4fe9-b1a2-7b29bde49b19 4558062d-b26b-4b7d-9adf-5ba876b07d08 276bcfc2-822c-4d05-88f0-be6c067a20c1 |
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 |
Universidade Federal de Santa Maria |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Bioquímica |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/11085/1/ALEDSONTORRES.pdf http://repositorio.ufsm.br/bitstream/1/11085/2/ALEDSONTORRES.pdf.txt http://repositorio.ufsm.br/bitstream/1/11085/3/ALEDSONTORRES.pdf.jpg |
bitstream.checksum.fl_str_mv |
f964591d9a022846aed4a3182d8f88a7 a6ff7b8900fd7cf8876e829bc9f4852d ce39cf9d37610fa334816b00d8529430 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
|
_version_ |
1794524290843934720 |