Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study

Detalhes bibliográficos
Autor(a) principal: Publio, B.C.
Data de Publicação: 2018
Outros Autores: Moura, T.A., Lima, C.H.M., Rocha, M.S.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: https://doi.org/10.1016/j.ijbiomac.2018.01.142
http://www.locus.ufv.br/handle/123456789/19129
Resumo: We have performed a biophysical characterization, at single molecule level, of the interaction between the DNA molecule and the biogenic polyamine putrescine. By using force spectroscopy, we were able to monitor the complexes formation as putrescine is added to the sample, determining the mechanical properties of such complexes and the physicochemical (binding) parameters of the interaction for three different ionic strengths. In particular, it was shown that the behavior of the equilibrium binding constant as a function of the counterion concentration deviates from the prediction of the Record-Lohman model. The measured constants were (1.3 ± 0.2) × 10^5 M^- 1 for [Na] = 150 mM, (2.1 ± 0.2) × 10^5 M^- 1 for [Na] = 10 mM, and (2.2 ± 0.3) × 10^5 M^- 1 for [Na] = 1 mM. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength. From these analysis, the DNA-putrescine binding mechanisms are inferred, and a comparison with results reported for ordinary bivalent ions like magnesium is performed. Such study provides new insights on the general behavior of the DNA interactions with biogenic polyamines.
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spelling Publio, B.C.Moura, T.A.Lima, C.H.M.Rocha, M.S.2018-04-25T15:30:41Z2018-04-25T15:30:41Z2018-02-2601418130https://doi.org/10.1016/j.ijbiomac.2018.01.142http://www.locus.ufv.br/handle/123456789/19129We have performed a biophysical characterization, at single molecule level, of the interaction between the DNA molecule and the biogenic polyamine putrescine. By using force spectroscopy, we were able to monitor the complexes formation as putrescine is added to the sample, determining the mechanical properties of such complexes and the physicochemical (binding) parameters of the interaction for three different ionic strengths. In particular, it was shown that the behavior of the equilibrium binding constant as a function of the counterion concentration deviates from the prediction of the Record-Lohman model. The measured constants were (1.3 ± 0.2) × 10^5 M^- 1 for [Na] = 150 mM, (2.1 ± 0.2) × 10^5 M^- 1 for [Na] = 10 mM, and (2.2 ± 0.3) × 10^5 M^- 1 for [Na] = 1 mM. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength. From these analysis, the DNA-putrescine binding mechanisms are inferred, and a comparison with results reported for ordinary bivalent ions like magnesium is performed. Such study provides new insights on the general behavior of the DNA interactions with biogenic polyamines.engInternational Journal of Biological Macromoleculesv. 112, p. 175-178, June 2018Elsevier B.Vinfo:eu-repo/semantics/openAccessPutrescineDNASingle molecule force spectroscopyBiophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule studyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALartigo.pdfartigo.pdftexto completoapplication/pdf1005039https://locus.ufv.br//bitstream/123456789/19129/1/artigo.pdf9977d27a9c94ade7912c237dafe526c9MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/19129/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52THUMBNAILartigo.pdf.jpgartigo.pdf.jpgIM Thumbnailimage/jpeg3671https://locus.ufv.br//bitstream/123456789/19129/3/artigo.pdf.jpg5a36b78455cd6a4e0b26ee9ead27167fMD53123456789/191292018-04-25 23:00:45.049oai:locus.ufv.br:123456789/19129Tk9URTogUExBQ0UgWU9VUiBPV04gTElDRU5TRSBIRVJFClRoaXMgc2FtcGxlIGxpY2Vuc2UgaXMgcHJvdmlkZWQgZm9yIGluZm9ybWF0aW9uYWwgcHVycG9zZXMgb25seS4KCk5PTi1FWENMVVNJVkUgRElTVFJJQlVUSU9OIExJQ0VOU0UKCkJ5IHNpZ25pbmcgYW5kIHN1Ym1pdHRpbmcgdGhpcyBsaWNlbnNlLCB5b3UgKHRoZSBhdXRob3Iocykgb3IgY29weXJpZ2h0Cm93bmVyKSBncmFudHMgdG8gRFNwYWNlIFVuaXZlcnNpdHkgKERTVSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBEU1UgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgRFNVIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgRFNVIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gRFNVLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpEU1Ugd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452018-04-26T02:00:45LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.en.fl_str_mv Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
title Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
spellingShingle Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
Publio, B.C.
Putrescine
DNA
Single molecule force spectroscopy
title_short Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
title_full Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
title_fullStr Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
title_full_unstemmed Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
title_sort Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: a single molecule study
author Publio, B.C.
author_facet Publio, B.C.
Moura, T.A.
Lima, C.H.M.
Rocha, M.S.
author_role author
author2 Moura, T.A.
Lima, C.H.M.
Rocha, M.S.
author2_role author
author
author
dc.contributor.author.fl_str_mv Publio, B.C.
Moura, T.A.
Lima, C.H.M.
Rocha, M.S.
dc.subject.pt-BR.fl_str_mv Putrescine
DNA
Single molecule force spectroscopy
topic Putrescine
DNA
Single molecule force spectroscopy
description We have performed a biophysical characterization, at single molecule level, of the interaction between the DNA molecule and the biogenic polyamine putrescine. By using force spectroscopy, we were able to monitor the complexes formation as putrescine is added to the sample, determining the mechanical properties of such complexes and the physicochemical (binding) parameters of the interaction for three different ionic strengths. In particular, it was shown that the behavior of the equilibrium binding constant as a function of the counterion concentration deviates from the prediction of the Record-Lohman model. The measured constants were (1.3 ± 0.2) × 10^5 M^- 1 for [Na] = 150 mM, (2.1 ± 0.2) × 10^5 M^- 1 for [Na] = 10 mM, and (2.2 ± 0.3) × 10^5 M^- 1 for [Na] = 1 mM. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength. From these analysis, the DNA-putrescine binding mechanisms are inferred, and a comparison with results reported for ordinary bivalent ions like magnesium is performed. Such study provides new insights on the general behavior of the DNA interactions with biogenic polyamines.
publishDate 2018
dc.date.accessioned.fl_str_mv 2018-04-25T15:30:41Z
dc.date.available.fl_str_mv 2018-04-25T15:30:41Z
dc.date.issued.fl_str_mv 2018-02-26
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 https://doi.org/10.1016/j.ijbiomac.2018.01.142
http://www.locus.ufv.br/handle/123456789/19129
dc.identifier.issn.none.fl_str_mv 01418130
identifier_str_mv 01418130
url https://doi.org/10.1016/j.ijbiomac.2018.01.142
http://www.locus.ufv.br/handle/123456789/19129
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartofseries.pt-BR.fl_str_mv v. 112, p. 175-178, June 2018
dc.rights.driver.fl_str_mv Elsevier B.V
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Elsevier B.V
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv International Journal of Biological Macromolecules
publisher.none.fl_str_mv International Journal of Biological Macromolecules
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
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