RPA-1 from Leishmania sp.: Recombinant Protein Expression and Purification, Molecular Modeling, and Molecular Dynamics Simulations Protocols

Registro completo de metadados
MetadadosDescriçãoIdioma
Autor(es): dc.contributorUniversidade Estadual Paulista (Unesp)-
Autor(es): dc.creatorFernandes, Carlos A. H. [UNESP]-
Autor(es): dc.creatorMorea, Edna G. O. [UNESP]-
Autor(es): dc.creatorCano, Maria Isabel N. [UNESP]-
Data de aceite: dc.date.accessioned2022-02-22T00:46:28Z-
Data de disponibilização: dc.date.available2022-02-22T00:46:28Z-
Data de envio: dc.date.issued2021-06-25-
Data de envio: dc.date.issued2021-06-25-
Data de envio: dc.date.issued2020-12-31-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.1007/978-1-0716-1290-3_10-
Fonte completa do material: dc.identifierhttp://hdl.handle.net/11449/206210-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/206210-
Descrição: dc.descriptionRPA is a conserved heterotrimeric complex and the major single-stranded DNA (ssDNA)-binding protein heterotrimeric complex, which in eukaryotes is formed by the RPA-1, RPA-2, and RPA-3 subunits. The main structural feature of RPA is the presence of the oligonucleotide/oligosaccharide-binding fold (OB-fold) domains, responsible for ssDNA binding and protein:protein interactions. Among the RPA subunits, RPA-1 bears three of the four OB folds involved with RPA-ssDNA binding, although in some organisms RPA-2 can also bind ssDNA. The OB-fold domains are also present in telomere end-binding proteins (TEBP), essential for chromosome end protection. RPA-1 from Leishmania sp., as well as RPA-1 from trypanosomatids, a group of early-divergent protozoa, shows some structural differences compared to higher eukaryote RPA-1. Also, RPA-1 from Leishmania sp., similar to TEBPs, may exert telomeric protective functions. Remarkably, different pieces of evidence have pointed out that trypanosomatids may not have OB fold-containing TEBPs. Moreover, recent data indicate that trypanosomatid RPA-1 may be considered a TEBP since it shares with TEBPs conserved functional and structural features. However, it is still unknown whether the RPA-1 protective telomeric role is exclusive to trypanosomatids or is also present in other primitive eukaryotes. Here, we describe a protocol to obtain highly purified and biologically active Leishmania amazonensis recombinant RPA-1, and to perform molecular modeling and molecular dynamics simulations methods which could be probably applied to functional and structural studies of homologous proteins in other primitive eukaryotes.-
Descrição: dc.descriptionDepartment of Biophysics and Pharmacology Biosciences Institute São Paulo State University (UNESP)-
Descrição: dc.descriptionDepartment of Chemical and Biological Sciences Biosciences Institute São Paulo State University (UNESP)-
Descrição: dc.descriptionDepartment of Biophysics and Pharmacology Biosciences Institute São Paulo State University (UNESP)-
Descrição: dc.descriptionDepartment of Chemical and Biological Sciences Biosciences Institute São Paulo State University (UNESP)-
Formato: dc.format169-191-
Idioma: dc.languageen-
Relação: dc.relationMethods in Molecular Biology-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectLeishmania-
Palavras-chave: dc.subjectOB-fold-
Palavras-chave: dc.subjectRPA-
Palavras-chave: dc.subjectTEBP-
Palavras-chave: dc.subjectTelomeres-
Palavras-chave: dc.subjectTrypanosomatids-
Título: dc.titleRPA-1 from Leishmania sp.: Recombinant Protein Expression and Purification, Molecular Modeling, and Molecular Dynamics Simulations Protocols-
Aparece nas coleções:Repositório Institucional - Unesp

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