Magnetic particles used in a new approach for designed protein crystallization

Bibliographic Details
Main Author: dos Santos, Raquel
Publication Date: 2021
Other Authors: Romão, Maria João, Roque, Ana Cecília A., Carvalho, Ana Luísa
Format: Article
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10362/141855
Summary: PD/BD/105753/2014 PTDC/BII-BIO/28878/2017 UIDB/04378/ 2020 POCI-01-0145-FEDER-007728
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spelling Magnetic particles used in a new approach for designed protein crystallizationChemistry(all)Materials Science(all)Condensed Matter PhysicsPD/BD/105753/2014 PTDC/BII-BIO/28878/2017 UIDB/04378/ 2020 POCI-01-0145-FEDER-007728After more than one hundred and thirty thousand protein structures determined by X-ray crystallography, the challenge of protein crystallization for 3D structure determination remains. In the quest for additives for efficient protein crystallization, inorganic materials emerge as an alternative. Magnetic particles (MPs) are versatile inorganic materials, easy to use, modify and manipulate in a wide range of biological assays. The potential of using functionalised MPs as crystallization chaperones for protein crystallization was shown in this work. MPs with distinct coatings were rationally designed to promote protein crystallization by affinity-triggered heterogeneous nucleation. Hen egg white lysozyme (HEWL) and trypsin, were crystallized in the presence of MPs either bare or coated with a polysaccharide (chitin) or a protein (casein), respectively. The addition of MPs was characterized in terms of bound protein to the MPs, crystal morphology, time-lapse of crystal emergence, crystallization yield fold change and crystal diffraction quality for structure determination. The MPs additives have shown to bind to the respective target protein, and to promote nucleation and crystal growth without compromising crystal morphology. On the other hand, MPs addition led to faster detectable crystal emergence and up to 13 times higher crystallization yield, addressing some the challenges in protein crystallization, the main bottleneck of macromolecular crystallography. Structure determination of the protein crystallized in the presence of MPs revealed that the structural characteristics of the protein remained unchanged, as shown by the superposition with PDB annotated proteins. Moreover, and unlike most reported cases, it was possible to exclude the inhibitor benzamidine during trypsin crystallisation, which is a remarkable result opening new prospects in enzyme engineering and drug design. Our results show that MPs coated with affinity ligands to target proteins can be used as controlled and tailor-made crystallization inducers.DQ - Departamento de QuímicaUCIBIO - Applied Molecular Biosciences UnitRUNdos Santos, RaquelRomão, Maria JoãoRoque, Ana Cecília A.Carvalho, Ana Luísa2022-07-13T22:23:03Z2021-02-072021-02-07T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article8application/pdfhttp://hdl.handle.net/10362/141855eng1466-8033PURE: 28431465https://doi.org/10.1039/d0ce01529finfo:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2024-05-22T18:03:27Zoai:run.unl.pt:10362/141855Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T17:34:15.852363Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Magnetic particles used in a new approach for designed protein crystallization
title Magnetic particles used in a new approach for designed protein crystallization
spellingShingle Magnetic particles used in a new approach for designed protein crystallization
dos Santos, Raquel
Chemistry(all)
Materials Science(all)
Condensed Matter Physics
title_short Magnetic particles used in a new approach for designed protein crystallization
title_full Magnetic particles used in a new approach for designed protein crystallization
title_fullStr Magnetic particles used in a new approach for designed protein crystallization
title_full_unstemmed Magnetic particles used in a new approach for designed protein crystallization
title_sort Magnetic particles used in a new approach for designed protein crystallization
author dos Santos, Raquel
author_facet dos Santos, Raquel
Romão, Maria João
Roque, Ana Cecília A.
Carvalho, Ana Luísa
author_role author
author2 Romão, Maria João
Roque, Ana Cecília A.
Carvalho, Ana Luísa
author2_role author
author
author
dc.contributor.none.fl_str_mv DQ - Departamento de Química
UCIBIO - Applied Molecular Biosciences Unit
RUN
dc.contributor.author.fl_str_mv dos Santos, Raquel
Romão, Maria João
Roque, Ana Cecília A.
Carvalho, Ana Luísa
dc.subject.por.fl_str_mv Chemistry(all)
Materials Science(all)
Condensed Matter Physics
topic Chemistry(all)
Materials Science(all)
Condensed Matter Physics
description PD/BD/105753/2014 PTDC/BII-BIO/28878/2017 UIDB/04378/ 2020 POCI-01-0145-FEDER-007728
publishDate 2021
dc.date.none.fl_str_mv 2021-02-07
2021-02-07T00:00:00Z
2022-07-13T22:23:03Z
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://hdl.handle.net/10362/141855
url http://hdl.handle.net/10362/141855
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1466-8033
PURE: 28431465
https://doi.org/10.1039/d0ce01529f
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 8
application/pdf
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron:RCAAP
instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron_str RCAAP
institution RCAAP
reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
collection Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
repository.name.fl_str_mv Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
repository.mail.fl_str_mv info@rcaap.pt
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