Measuring transverse relaxation in highly paramagnetic systems

Detalhes bibliográficos
Autor(a) principal: Invernici, Michele
Data de Publicação: 2020
Outros Autores: Trindade, Inês B., Cantini, Francesca, Louro, Ricardo O., Piccioli, Mario
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/120780
Resumo: The enhancement of nuclear relaxation rates due to the interaction with a paramagnetic center (known as Paramagnetic Relaxation Enhancement) is a powerful source of structural and dynamics information, widely used in structural biology. However, many signals affected by the hyperfine interaction relax faster than the evolution periods of common NMR experiments and therefore they are broadened beyond detection. This gives rise to a so-called blind sphere around the paramagnetic center, which is a major limitation in the use of PREs. Reducing the blind sphere is extremely important in paramagnetic metalloproteins. The identification, characterization, and proper structural restraining of the first coordination sphere of the metal ion(s) and its immediate neighboring regions is key to understand their biological function. The novel HSQC scheme we propose here, that we termed R2-weighted, HSQC-AP, achieves this aim by detecting signals that escaped detection in a conventional HSQC experiment and provides fully reliable R2 values in the range of 1H R2 rates ca. 50–400 s−1. Independently on the type of paramagnetic center and on the size of the molecule, this experiment decreases the radius of the blind sphere and increases the number of detectable PREs. Here, we report the validation of this approach for the case of PioC, a small protein containing a high potential 4Fe-4S cluster in the reduced [Fe4S4]2+ form. The blind sphere was contracted to a minimal extent, enabling the measurement of R2 rates for the cluster coordinating residues.
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spelling Measuring transverse relaxation in highly paramagnetic systemsIron sulfur proteinsNMR based structural restraintsParamagnetic NMRParamagnetic relaxation enhancementPulse sequencesTransverse relaxationBiochemistrySpectroscopyThe enhancement of nuclear relaxation rates due to the interaction with a paramagnetic center (known as Paramagnetic Relaxation Enhancement) is a powerful source of structural and dynamics information, widely used in structural biology. However, many signals affected by the hyperfine interaction relax faster than the evolution periods of common NMR experiments and therefore they are broadened beyond detection. This gives rise to a so-called blind sphere around the paramagnetic center, which is a major limitation in the use of PREs. Reducing the blind sphere is extremely important in paramagnetic metalloproteins. The identification, characterization, and proper structural restraining of the first coordination sphere of the metal ion(s) and its immediate neighboring regions is key to understand their biological function. The novel HSQC scheme we propose here, that we termed R2-weighted, HSQC-AP, achieves this aim by detecting signals that escaped detection in a conventional HSQC experiment and provides fully reliable R2 values in the range of 1H R2 rates ca. 50–400 s−1. Independently on the type of paramagnetic center and on the size of the molecule, this experiment decreases the radius of the blind sphere and increases the number of detectable PREs. Here, we report the validation of this approach for the case of PioC, a small protein containing a high potential 4Fe-4S cluster in the reduced [Fe4S4]2+ form. The blind sphere was contracted to a minimal extent, enabling the measurement of R2 rates for the cluster coordinating residues.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNInvernici, MicheleTrindade, Inês B.Cantini, FrancescaLouro, Ricardo O.Piccioli, Mario2021-07-09T22:17:14Z2020-09-012020-09-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttp://hdl.handle.net/10362/120780eng0925-2738PURE: 28003447https://doi.org/10.1007/s10858-020-00334-winfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:03:14Zoai:run.unl.pt:10362/120780Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:44:26.671071Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Measuring transverse relaxation in highly paramagnetic systems
title Measuring transverse relaxation in highly paramagnetic systems
spellingShingle Measuring transverse relaxation in highly paramagnetic systems
Invernici, Michele
Iron sulfur proteins
NMR based structural restraints
Paramagnetic NMR
Paramagnetic relaxation enhancement
Pulse sequences
Transverse relaxation
Biochemistry
Spectroscopy
title_short Measuring transverse relaxation in highly paramagnetic systems
title_full Measuring transverse relaxation in highly paramagnetic systems
title_fullStr Measuring transverse relaxation in highly paramagnetic systems
title_full_unstemmed Measuring transverse relaxation in highly paramagnetic systems
title_sort Measuring transverse relaxation in highly paramagnetic systems
author Invernici, Michele
author_facet Invernici, Michele
Trindade, Inês B.
Cantini, Francesca
Louro, Ricardo O.
Piccioli, Mario
author_role author
author2 Trindade, Inês B.
Cantini, Francesca
Louro, Ricardo O.
Piccioli, Mario
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv Invernici, Michele
Trindade, Inês B.
Cantini, Francesca
Louro, Ricardo O.
Piccioli, Mario
dc.subject.por.fl_str_mv Iron sulfur proteins
NMR based structural restraints
Paramagnetic NMR
Paramagnetic relaxation enhancement
Pulse sequences
Transverse relaxation
Biochemistry
Spectroscopy
topic Iron sulfur proteins
NMR based structural restraints
Paramagnetic NMR
Paramagnetic relaxation enhancement
Pulse sequences
Transverse relaxation
Biochemistry
Spectroscopy
description The enhancement of nuclear relaxation rates due to the interaction with a paramagnetic center (known as Paramagnetic Relaxation Enhancement) is a powerful source of structural and dynamics information, widely used in structural biology. However, many signals affected by the hyperfine interaction relax faster than the evolution periods of common NMR experiments and therefore they are broadened beyond detection. This gives rise to a so-called blind sphere around the paramagnetic center, which is a major limitation in the use of PREs. Reducing the blind sphere is extremely important in paramagnetic metalloproteins. The identification, characterization, and proper structural restraining of the first coordination sphere of the metal ion(s) and its immediate neighboring regions is key to understand their biological function. The novel HSQC scheme we propose here, that we termed R2-weighted, HSQC-AP, achieves this aim by detecting signals that escaped detection in a conventional HSQC experiment and provides fully reliable R2 values in the range of 1H R2 rates ca. 50–400 s−1. Independently on the type of paramagnetic center and on the size of the molecule, this experiment decreases the radius of the blind sphere and increases the number of detectable PREs. Here, we report the validation of this approach for the case of PioC, a small protein containing a high potential 4Fe-4S cluster in the reduced [Fe4S4]2+ form. The blind sphere was contracted to a minimal extent, enabling the measurement of R2 rates for the cluster coordinating residues.
publishDate 2020
dc.date.none.fl_str_mv 2020-09-01
2020-09-01T00:00:00Z
2021-07-09T22:17:14Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/120780
url http://hdl.handle.net/10362/120780
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0925-2738
PURE: 28003447
https://doi.org/10.1007/s10858-020-00334-w
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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