Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s00249-016-1121-6 http://hdl.handle.net/11449/161994 |
Resumo: | Oxy-HbRa thermal stability was evaluated by dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS) at pH 5.0, 7.0, 8.0, and 9.0. DLS results show that oxy-HbRa, at pH 7.0 and 5.0, remains stable up to 56 degrees C, undergoing denaturation/aggregation in acidic media above 60 degrees C, followed by partial sedimentation of aggregates. At alkaline pH values 8.0 and 9.0, oxy-HbRa oligomeric dissociation is observed above 30 degrees C, before denaturation. SAXS data show that oxy-HbRa, at 20 degrees C, is in its native form, displaying radius of gyration (R-g) and particle maximum dimension (D-max) of 108 +/- 1 and 300 +/- 10 angstrom, respectively. Oxy-HbRa, at pH 7.0, undergoes denaturation/aggregation at 60 degrees C. At pH 5.0-6.0, HbRa thermal denaturation/aggregation start earlier, at 50 degrees C, accompanied by an increase of R-g and D-max values. However, an overlap of oligomeric dissociation and denaturation in the system is observed upon temperature increase, with an increase in R-g and D-max. Analysis of experimental p(r) curves as a linear combination of theoretical curves obtained for HbGp fragments from the crystal structure shows an increasing contribution of dodecamer (abcd)(3) and tetramer (abcd) in solution, as a function of pH values (8.0 and 9.0) and temperature. Finally, our data show, for the first time, that oxy-HbRa, in neutral and acidic media, does not undergo oligomeric dissociation before denaturation, while in alkaline media the oligomeric dissociation process is an important step in the thermal denaturation. |
id |
UNSP_2a96c0aaf2659accef2532b18c3015df |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/161994 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studiesErythrocruorinsThermal stabilityDenaturation/aggregationOligomeric dissociationpHOxy-HbRa thermal stability was evaluated by dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS) at pH 5.0, 7.0, 8.0, and 9.0. DLS results show that oxy-HbRa, at pH 7.0 and 5.0, remains stable up to 56 degrees C, undergoing denaturation/aggregation in acidic media above 60 degrees C, followed by partial sedimentation of aggregates. At alkaline pH values 8.0 and 9.0, oxy-HbRa oligomeric dissociation is observed above 30 degrees C, before denaturation. SAXS data show that oxy-HbRa, at 20 degrees C, is in its native form, displaying radius of gyration (R-g) and particle maximum dimension (D-max) of 108 +/- 1 and 300 +/- 10 angstrom, respectively. Oxy-HbRa, at pH 7.0, undergoes denaturation/aggregation at 60 degrees C. At pH 5.0-6.0, HbRa thermal denaturation/aggregation start earlier, at 50 degrees C, accompanied by an increase of R-g and D-max values. However, an overlap of oligomeric dissociation and denaturation in the system is observed upon temperature increase, with an increase in R-g and D-max. Analysis of experimental p(r) curves as a linear combination of theoretical curves obtained for HbGp fragments from the crystal structure shows an increasing contribution of dodecamer (abcd)(3) and tetramer (abcd) in solution, as a function of pH values (8.0 and 9.0) and temperature. Finally, our data show, for the first time, that oxy-HbRa, in neutral and acidic media, does not undergo oligomeric dissociation before denaturation, while in alkaline media the oligomeric dissociation process is an important step in the thermal denaturation.Inst Quim Sao Carlos, Sao Carlos, SP, BrazilUniv Estado Mato Grosso, Barra Do Bugres, MT, BrazilUniv Estadual Paulista, Registro, SP, BrazilUniv Estadual Paulista, Registro, SP, BrazilSpringerInst Quim Sao CarlosUniv Estado Mato GrossoUniversidade Estadual Paulista (Unesp)Carvalho, Jose Wilson P.Carvalho, Francisco A. O.Santiago, Patricia S. [UNESP]Tabak, Marcel2018-11-26T17:06:27Z2018-11-26T17:06:27Z2016-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article549-563application/pdfhttp://dx.doi.org/10.1007/s00249-016-1121-6European Biophysics Journal With Biophysics Letters. New York: Springer, v. 45, n. 6, p. 549-563, 2016.0175-7571http://hdl.handle.net/11449/16199410.1007/s00249-016-1121-6WOS:000384722500006WOS000384722500006.pdf67053670106620870000-0002-6205-9441Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengEuropean Biophysics Journal With Biophysics Letters0,604info:eu-repo/semantics/openAccess2024-05-03T13:19:57Zoai:repositorio.unesp.br:11449/161994Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-05-03T13:19:57Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
title |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
spellingShingle |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies Carvalho, Jose Wilson P. Erythrocruorins Thermal stability Denaturation/aggregation Oligomeric dissociation pH |
title_short |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
title_full |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
title_fullStr |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
title_full_unstemmed |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
title_sort |
Thermal stability of extracellular hemoglobin of Rhinodrilus alatus (HbRa): DLS and SAXS studies |
author |
Carvalho, Jose Wilson P. |
author_facet |
Carvalho, Jose Wilson P. Carvalho, Francisco A. O. Santiago, Patricia S. [UNESP] Tabak, Marcel |
author_role |
author |
author2 |
Carvalho, Francisco A. O. Santiago, Patricia S. [UNESP] Tabak, Marcel |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Inst Quim Sao Carlos Univ Estado Mato Grosso Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Carvalho, Jose Wilson P. Carvalho, Francisco A. O. Santiago, Patricia S. [UNESP] Tabak, Marcel |
dc.subject.por.fl_str_mv |
Erythrocruorins Thermal stability Denaturation/aggregation Oligomeric dissociation pH |
topic |
Erythrocruorins Thermal stability Denaturation/aggregation Oligomeric dissociation pH |
description |
Oxy-HbRa thermal stability was evaluated by dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS) at pH 5.0, 7.0, 8.0, and 9.0. DLS results show that oxy-HbRa, at pH 7.0 and 5.0, remains stable up to 56 degrees C, undergoing denaturation/aggregation in acidic media above 60 degrees C, followed by partial sedimentation of aggregates. At alkaline pH values 8.0 and 9.0, oxy-HbRa oligomeric dissociation is observed above 30 degrees C, before denaturation. SAXS data show that oxy-HbRa, at 20 degrees C, is in its native form, displaying radius of gyration (R-g) and particle maximum dimension (D-max) of 108 +/- 1 and 300 +/- 10 angstrom, respectively. Oxy-HbRa, at pH 7.0, undergoes denaturation/aggregation at 60 degrees C. At pH 5.0-6.0, HbRa thermal denaturation/aggregation start earlier, at 50 degrees C, accompanied by an increase of R-g and D-max values. However, an overlap of oligomeric dissociation and denaturation in the system is observed upon temperature increase, with an increase in R-g and D-max. Analysis of experimental p(r) curves as a linear combination of theoretical curves obtained for HbGp fragments from the crystal structure shows an increasing contribution of dodecamer (abcd)(3) and tetramer (abcd) in solution, as a function of pH values (8.0 and 9.0) and temperature. Finally, our data show, for the first time, that oxy-HbRa, in neutral and acidic media, does not undergo oligomeric dissociation before denaturation, while in alkaline media the oligomeric dissociation process is an important step in the thermal denaturation. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-09-01 2018-11-26T17:06:27Z 2018-11-26T17:06:27Z |
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/10.1007/s00249-016-1121-6 European Biophysics Journal With Biophysics Letters. New York: Springer, v. 45, n. 6, p. 549-563, 2016. 0175-7571 http://hdl.handle.net/11449/161994 10.1007/s00249-016-1121-6 WOS:000384722500006 WOS000384722500006.pdf 6705367010662087 0000-0002-6205-9441 |
url |
http://dx.doi.org/10.1007/s00249-016-1121-6 http://hdl.handle.net/11449/161994 |
identifier_str_mv |
European Biophysics Journal With Biophysics Letters. New York: Springer, v. 45, n. 6, p. 549-563, 2016. 0175-7571 10.1007/s00249-016-1121-6 WOS:000384722500006 WOS000384722500006.pdf 6705367010662087 0000-0002-6205-9441 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
European Biophysics Journal With Biophysics Letters 0,604 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
549-563 application/pdf |
dc.publisher.none.fl_str_mv |
Springer |
publisher.none.fl_str_mv |
Springer |
dc.source.none.fl_str_mv |
Web of Science 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_ |
1799964669433085952 |