New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution
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Publication Date: | 2016 |
Other Authors: | , , , , , , , |
Format: | Article |
Language: | eng |
Source: | Repositório Institucional da UNESP |
Download full: | http://dx.doi.org/10.1038/srep29976 http://hdl.handle.net/11449/161741 |
Summary: | Among all proteins localized in the Golgi apparatus, a two-PDZ (PSD95/DlgA/Zo-1) domain protein plays an important role in the assembly of the cisternae. This Golgi Reassembly and Stacking Protein (GRASP) has puzzled researchers due to its large array of functions and relevance in Golgi functionality. We report here a biochemical and biophysical study of the GRASP55/65 homologue in Cryptococcus neoformans (CnGRASP). Bioinformatic analysis, static fluorescence and circular dichroism spectroscopies, calorimetry, small angle X-ray scattering, solution nuclear magnetic resonance, size exclusion chromatography and proteolysis assays were used to unravel structural features of the full-length CnGRASP. We detected the coexistence of regular secondary structures and large amounts of disordered regions. The overall structure is less compact than a regular globular protein and the high structural flexibility makes its hydrophobic core more accessible to solvent. Our results indicate an unusual behavior of CnGRASP in solution, closely resembling a class of intrinsically disordered proteins called molten globule proteins. To the best of our knowledge, this is the first structural characterization of a full-length GRASP and observation of a molten globule-like behavior in the GRASP family. The possible implications of this and how it could explain the multiple facets of this intriguing class of proteins are discussed. |
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New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solutionAmong all proteins localized in the Golgi apparatus, a two-PDZ (PSD95/DlgA/Zo-1) domain protein plays an important role in the assembly of the cisternae. This Golgi Reassembly and Stacking Protein (GRASP) has puzzled researchers due to its large array of functions and relevance in Golgi functionality. We report here a biochemical and biophysical study of the GRASP55/65 homologue in Cryptococcus neoformans (CnGRASP). Bioinformatic analysis, static fluorescence and circular dichroism spectroscopies, calorimetry, small angle X-ray scattering, solution nuclear magnetic resonance, size exclusion chromatography and proteolysis assays were used to unravel structural features of the full-length CnGRASP. We detected the coexistence of regular secondary structures and large amounts of disordered regions. The overall structure is less compact than a regular globular protein and the high structural flexibility makes its hydrophobic core more accessible to solvent. Our results indicate an unusual behavior of CnGRASP in solution, closely resembling a class of intrinsically disordered proteins called molten globule proteins. To the best of our knowledge, this is the first structural characterization of a full-length GRASP and observation of a molten globule-like behavior in the GRASP family. The possible implications of this and how it could explain the multiple facets of this intriguing class of proteins are discussed.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Wellcome Trust (UK)Instituto Nacional de Ciencia e Tecnologia de Inovacao em Doencas Negligenciadas (INCT-IDN)Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Fis, Lab Biofis Mol, Ribeirao Preto, SP, BrazilUniv Sao Paulo, Inst Fis Sao Carlos, Dept Fis & Informat, Sao Carlos, SP, BrazilUniv Estadual Paulista Julio Mesquita, Ctr Multiusuario Inovacao Biomol, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, Sao Jose Do Rio Preto, BrazilUniv Fed Rio Grande do Sul, Ctr Biotecnol, Porto Alegre, RS, BrazilFundacao Oswaldo Cruz, FIOCRUZ, CDTS, Rio De Janeiro, BrazilUniv Fed Rio de Janeiro, Inst Microbiol Paulo Goes, Rio De Janeiro, BrazilUniv Estadual Paulista Julio Mesquita, Ctr Multiusuario Inovacao Biomol, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, Sao Jose Do Rio Preto, BrazilFAPESP: 2012/20367-3FAPESP: 2012/13309-7CNPq: 308380/2013-4Nature Publishing GroupUniversidade de São Paulo (USP)Universidade Estadual Paulista (Unesp)Univ Fed Rio Grande do SulFundacao Oswaldo CruzUniversidade Federal do Rio de Janeiro (UFRJ)Mendes, Luis F. S.Garcia, Assuero F.Kumagai, Patricia S.Morais, Fabio R. de [UNESP]Melo, Fernando A. [UNESP]Kmetzsch, LiviaVainstein, Marilene H.Rodrigues, Marcio L.Costa-Filho, Antonio J.2018-11-26T16:48:26Z2018-11-26T16:48:26Z2016-07-20info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article14application/pdfhttp://dx.doi.org/10.1038/srep29976Scientific Reports. London: Nature Publishing Group, v. 6, 14 p., 2016.2045-2322http://hdl.handle.net/11449/16174110.1038/srep29976WOS:000379978600001WOS000379978600001.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengScientific Reports1,533info:eu-repo/semantics/openAccess2024-01-04T06:27:10Zoai:repositorio.unesp.br:11449/161741Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-01-04T06:27:10Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
title |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
spellingShingle |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution Mendes, Luis F. S. |
title_short |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
title_full |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
title_fullStr |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
title_full_unstemmed |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
title_sort |
New structural insights into Golgi Reassembly and Stacking Protein (GRASP) in solution |
author |
Mendes, Luis F. S. |
author_facet |
Mendes, Luis F. S. Garcia, Assuero F. Kumagai, Patricia S. Morais, Fabio R. de [UNESP] Melo, Fernando A. [UNESP] Kmetzsch, Livia Vainstein, Marilene H. Rodrigues, Marcio L. Costa-Filho, Antonio J. |
author_role |
author |
author2 |
Garcia, Assuero F. Kumagai, Patricia S. Morais, Fabio R. de [UNESP] Melo, Fernando A. [UNESP] Kmetzsch, Livia Vainstein, Marilene H. Rodrigues, Marcio L. Costa-Filho, Antonio J. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Estadual Paulista (Unesp) Univ Fed Rio Grande do Sul Fundacao Oswaldo Cruz Universidade Federal do Rio de Janeiro (UFRJ) |
dc.contributor.author.fl_str_mv |
Mendes, Luis F. S. Garcia, Assuero F. Kumagai, Patricia S. Morais, Fabio R. de [UNESP] Melo, Fernando A. [UNESP] Kmetzsch, Livia Vainstein, Marilene H. Rodrigues, Marcio L. Costa-Filho, Antonio J. |
description |
Among all proteins localized in the Golgi apparatus, a two-PDZ (PSD95/DlgA/Zo-1) domain protein plays an important role in the assembly of the cisternae. This Golgi Reassembly and Stacking Protein (GRASP) has puzzled researchers due to its large array of functions and relevance in Golgi functionality. We report here a biochemical and biophysical study of the GRASP55/65 homologue in Cryptococcus neoformans (CnGRASP). Bioinformatic analysis, static fluorescence and circular dichroism spectroscopies, calorimetry, small angle X-ray scattering, solution nuclear magnetic resonance, size exclusion chromatography and proteolysis assays were used to unravel structural features of the full-length CnGRASP. We detected the coexistence of regular secondary structures and large amounts of disordered regions. The overall structure is less compact than a regular globular protein and the high structural flexibility makes its hydrophobic core more accessible to solvent. Our results indicate an unusual behavior of CnGRASP in solution, closely resembling a class of intrinsically disordered proteins called molten globule proteins. To the best of our knowledge, this is the first structural characterization of a full-length GRASP and observation of a molten globule-like behavior in the GRASP family. The possible implications of this and how it could explain the multiple facets of this intriguing class of proteins are discussed. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-07-20 2018-11-26T16:48:26Z 2018-11-26T16:48:26Z |
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/10.1038/srep29976 Scientific Reports. London: Nature Publishing Group, v. 6, 14 p., 2016. 2045-2322 http://hdl.handle.net/11449/161741 10.1038/srep29976 WOS:000379978600001 WOS000379978600001.pdf |
url |
http://dx.doi.org/10.1038/srep29976 http://hdl.handle.net/11449/161741 |
identifier_str_mv |
Scientific Reports. London: Nature Publishing Group, v. 6, 14 p., 2016. 2045-2322 10.1038/srep29976 WOS:000379978600001 WOS000379978600001.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Scientific Reports 1,533 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
14 application/pdf |
dc.publisher.none.fl_str_mv |
Nature Publishing Group |
publisher.none.fl_str_mv |
Nature Publishing Group |
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 |
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