Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Genetics and Molecular Biology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000300416 |
Resumo: | Abstract Carbonic anhydrases (CA) belong to the family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate. In the present work, we characterized the cDNAs of four Paracoccidioides CAs (CA1, CA2, CA3, and CA4). In the presence of CO2, there was not a significant increase in fungal ca1, ca2 and ca4 gene expression. The ca1 transcript was induced during the mycelium-to-yeast transition, while ca2 and ca4 gene expression was much higher in yeast cells, when compared to mycelium and mycelium-to-yeast transition. The ca1 transcript was induced in yeast cells recovered directly from liver and spleen of infected mice, while transcripts for ca2 and ca4 were down-regulated. Recombinant CA1 (rCA1) and CA4 (rCA4), with 33 kDa and 32 kDa respectively, were obtained from bacteria. The enzymes rCA1 (β-class) and rCA4 (α-class) were characterized regarding pH, temperature, ions and amino acids addition influence. Both enzymes were stable at pHs 7.5-8.5 and temperatures of 30-35 °C. The enzymes were dramatically inhibited by Hg+2 and activated by Zn+2, while only rCA4 was stimulated by Fe2+. Among the amino acids tested (all in L configuration), arginine, lysine, tryptophan and histidine enhanced residual activity of rCA1 and rCA4. |
id |
SBG-1_957ea733b4cc6b10c065b7e968d77bd9 |
---|---|
oai_identifier_str |
oai:scielo:S1415-47572016000300416 |
network_acronym_str |
SBG-1 |
network_name_str |
Genetics and Molecular Biology |
repository_id_str |
|
spelling |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioidescarbonic anhydrasesParacoccidioidesCO2gene expressionenzyme assayAbstract Carbonic anhydrases (CA) belong to the family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate. In the present work, we characterized the cDNAs of four Paracoccidioides CAs (CA1, CA2, CA3, and CA4). In the presence of CO2, there was not a significant increase in fungal ca1, ca2 and ca4 gene expression. The ca1 transcript was induced during the mycelium-to-yeast transition, while ca2 and ca4 gene expression was much higher in yeast cells, when compared to mycelium and mycelium-to-yeast transition. The ca1 transcript was induced in yeast cells recovered directly from liver and spleen of infected mice, while transcripts for ca2 and ca4 were down-regulated. Recombinant CA1 (rCA1) and CA4 (rCA4), with 33 kDa and 32 kDa respectively, were obtained from bacteria. The enzymes rCA1 (β-class) and rCA4 (α-class) were characterized regarding pH, temperature, ions and amino acids addition influence. Both enzymes were stable at pHs 7.5-8.5 and temperatures of 30-35 °C. The enzymes were dramatically inhibited by Hg+2 and activated by Zn+2, while only rCA4 was stimulated by Fe2+. Among the amino acids tested (all in L configuration), arginine, lysine, tryptophan and histidine enhanced residual activity of rCA1 and rCA4.Sociedade Brasileira de Genética2016-09-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000300416Genetics and Molecular Biology v.39 n.3 2016reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/1678-4685-GMB-2015-0213info:eu-repo/semantics/openAccessTomazett,Mariana VieiraZanoelo,Fabiana FonsecaBailão,Elisa Flávia CardosoBailão,Alexandre MeloBorges,Clayton LuizSoares,Célia Maria de Almeidaeng2016-08-25T00:00:00Zoai:scielo:S1415-47572016000300416Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2016-08-25T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false |
dc.title.none.fl_str_mv |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
title |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
spellingShingle |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides Tomazett,Mariana Vieira carbonic anhydrases Paracoccidioides CO2 gene expression enzyme assay |
title_short |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
title_full |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
title_fullStr |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
title_full_unstemmed |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
title_sort |
Molecular and biochemical characterization of carbonic anhydrases of Paracoccidioides |
author |
Tomazett,Mariana Vieira |
author_facet |
Tomazett,Mariana Vieira Zanoelo,Fabiana Fonseca Bailão,Elisa Flávia Cardoso Bailão,Alexandre Melo Borges,Clayton Luiz Soares,Célia Maria de Almeida |
author_role |
author |
author2 |
Zanoelo,Fabiana Fonseca Bailão,Elisa Flávia Cardoso Bailão,Alexandre Melo Borges,Clayton Luiz Soares,Célia Maria de Almeida |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Tomazett,Mariana Vieira Zanoelo,Fabiana Fonseca Bailão,Elisa Flávia Cardoso Bailão,Alexandre Melo Borges,Clayton Luiz Soares,Célia Maria de Almeida |
dc.subject.por.fl_str_mv |
carbonic anhydrases Paracoccidioides CO2 gene expression enzyme assay |
topic |
carbonic anhydrases Paracoccidioides CO2 gene expression enzyme assay |
description |
Abstract Carbonic anhydrases (CA) belong to the family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate. In the present work, we characterized the cDNAs of four Paracoccidioides CAs (CA1, CA2, CA3, and CA4). In the presence of CO2, there was not a significant increase in fungal ca1, ca2 and ca4 gene expression. The ca1 transcript was induced during the mycelium-to-yeast transition, while ca2 and ca4 gene expression was much higher in yeast cells, when compared to mycelium and mycelium-to-yeast transition. The ca1 transcript was induced in yeast cells recovered directly from liver and spleen of infected mice, while transcripts for ca2 and ca4 were down-regulated. Recombinant CA1 (rCA1) and CA4 (rCA4), with 33 kDa and 32 kDa respectively, were obtained from bacteria. The enzymes rCA1 (β-class) and rCA4 (α-class) were characterized regarding pH, temperature, ions and amino acids addition influence. Both enzymes were stable at pHs 7.5-8.5 and temperatures of 30-35 °C. The enzymes were dramatically inhibited by Hg+2 and activated by Zn+2, while only rCA4 was stimulated by Fe2+. Among the amino acids tested (all in L configuration), arginine, lysine, tryptophan and histidine enhanced residual activity of rCA1 and rCA4. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-09-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000300416 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000300416 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/1678-4685-GMB-2015-0213 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
dc.source.none.fl_str_mv |
Genetics and Molecular Biology v.39 n.3 2016 reponame:Genetics and Molecular Biology instname:Sociedade Brasileira de Genética (SBG) instacron:SBG |
instname_str |
Sociedade Brasileira de Genética (SBG) |
instacron_str |
SBG |
institution |
SBG |
reponame_str |
Genetics and Molecular Biology |
collection |
Genetics and Molecular Biology |
repository.name.fl_str_mv |
Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG) |
repository.mail.fl_str_mv |
||editor@gmb.org.br |
_version_ |
1752122387064684544 |