Characterization and oxygen binding properties of des-Arg human hemoglobin
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1590/S0100-879X2009000600004 http://hdl.handle.net/11449/21668 |
Resumo: | The role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function. |
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Characterization and oxygen binding properties of des-Arg human hemoglobindes-Arg hemoglobinOxygen affinityThe role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNESP, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, BR-15054000 Sao Jose do Rio Preto, SP, BrazilUNESP, Inst Biociencias Letras & Ciencias Exatas, Dept Biol, BR-15054000 Sao Jose do Rio Preto, SP, BrazilUNESP, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, BR-15054000 Sao Jose do Rio Preto, SP, BrazilUNESP, Inst Biociencias Letras & Ciencias Exatas, Dept Biol, BR-15054000 Sao Jose do Rio Preto, SP, BrazilAssociação Brasileira de Divulgação Científica (ABRADIC)Universidade Estadual Paulista (Unesp)Tosqui, P. [UNESP]Bonini-Domingos, C.R. [UNESP]Colombo, M.F. [UNESP]2014-05-20T14:01:22Z2014-05-20T14:01:22Z2009-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article494-500application/pdfhttp://dx.doi.org/10.1590/S0100-879X2009000600004Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 42, n. 6, p. 494-500, 2009.0100-879Xhttp://hdl.handle.net/11449/2166810.1590/S0100-879X2009000600004S0100-879X2009000600004WOS:000266135800011S0100-879X2009000600004.pdf342581720964605432794280661767190000-0002-4603-9467SciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBrazilian Journal of Medical and Biological Research1.492info:eu-repo/semantics/openAccess2023-11-14T06:12:00Zoai:repositorio.unesp.br:11449/21668Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:39:36.704386Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
title |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
spellingShingle |
Characterization and oxygen binding properties of des-Arg human hemoglobin Tosqui, P. [UNESP] des-Arg hemoglobin Oxygen affinity |
title_short |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
title_full |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
title_fullStr |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
title_full_unstemmed |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
title_sort |
Characterization and oxygen binding properties of des-Arg human hemoglobin |
author |
Tosqui, P. [UNESP] |
author_facet |
Tosqui, P. [UNESP] Bonini-Domingos, C.R. [UNESP] Colombo, M.F. [UNESP] |
author_role |
author |
author2 |
Bonini-Domingos, C.R. [UNESP] Colombo, M.F. [UNESP] |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Tosqui, P. [UNESP] Bonini-Domingos, C.R. [UNESP] Colombo, M.F. [UNESP] |
dc.subject.por.fl_str_mv |
des-Arg hemoglobin Oxygen affinity |
topic |
des-Arg hemoglobin Oxygen affinity |
description |
The role of chloride in the stabilization of the deoxy conformation of hemoglobin (Hb), the low oxygen affinity state, has been studied in order to identify the nature of this binding. Previous studies have shown that arginines 141α could be involved in the binding of this ion to the protein. Thus, des-Arg Hb, human hemoglobin modified by removal of the α-chain C-terminal residue Arg141α, is a possible model for studies of these interactions. The loss of Arg141α and all the salt bridges in which it participates is associated with subtle structural perturbations of the α-chains, which include an increase in the conformational flexibility and further shift to the oxy state, increasing oxygen affinity. Thus, this Hb has been the target of many studies of structural and functional behavior along with medical applications. In the present study, we describe the biochemical characterization of des-Arg Hb by electrophoresis, high-performance liquid chromatography and mass spectroscopy. The effects of chloride binding on the oxygen affinity and on the cooperativity to des-Arg Hb and to native human hemoglobin, HbA, were measured and compared. We confirm that des-Arg Hb presents high oxygen affinity and low cooperativity in the presence of bound chloride and show that the binding of chloride to des-Arg does not change its functional characteristics as observed with HbA. These results indicate that Arg141α may be involved in the chloride effect on Hb oxygenation. Moreover, they show that these residues contribute to lower Hb oxygen affinity to a level compatible with its biological function. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-06-01 2014-05-20T14:01:22Z 2014-05-20T14:01:22Z |
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/10.1590/S0100-879X2009000600004 Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 42, n. 6, p. 494-500, 2009. 0100-879X http://hdl.handle.net/11449/21668 10.1590/S0100-879X2009000600004 S0100-879X2009000600004 WOS:000266135800011 S0100-879X2009000600004.pdf 3425817209646054 3279428066176719 0000-0002-4603-9467 |
url |
http://dx.doi.org/10.1590/S0100-879X2009000600004 http://hdl.handle.net/11449/21668 |
identifier_str_mv |
Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 42, n. 6, p. 494-500, 2009. 0100-879X 10.1590/S0100-879X2009000600004 S0100-879X2009000600004 WOS:000266135800011 S0100-879X2009000600004.pdf 3425817209646054 3279428066176719 0000-0002-4603-9467 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Brazilian Journal of Medical and Biological Research 1.492 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
494-500 application/pdf |
dc.publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica (ABRADIC) |
publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica (ABRADIC) |
dc.source.none.fl_str_mv |
SciELO 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 |
|
_version_ |
1808128840607727616 |