Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins

105 p.

Detalhes bibliográficos
Autor: Torcal Embeita, Bruno
Formato: tesis doctoral
Fecha de publicación:2016
País:España
Recursos:Universidad del País Vasco
Repositorio:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/20519
Acesso em linha:http://hdl.handle.net/10810/20519
Access Level:acceso abierto
Palavra-chave:molecular spectroscopy
photochemistry
quantum theory
espectroscopia molecular
fotoquímica
teoría cuántica
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spelling Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteinsTorcal Embeita, Brunomolecular spectroscopyphotochemistryquantum theoryespectroscopia molecularfotoquímicateoría cuántica105 p.Reversibly photoswitchable uorescent proteins (RSFPs) are genetically engineered proteins that can be switched by light absorption between a uorescent ON state and a dark OFF state. Among other applications they allow to increase the resolution beyond the difraction limit in cell imaging by uorescence microscopy. RSFPs have extended the possibilities of uorescence microscopy and other biotechnological tools, but the development of their properties is still far from being rationally designed. Thus, there might be much room for improvement if we manage to understand the switching mechanisms. The switching mechanisms in several negative RSFPs is being elucidated but still under debate. In this thesis I extend the theoretical knowledge about the photoswitching of negative RSFPs by studying the excited-state potential energy surface of both the ON- and the OFF-state. I compare three RSFPs, namely IrisFP, Dronpa and a fast switching single mutant of Dronpa called Dronpa2 to search for the origin of their diferent switching quantum yields in the ON-state. For the OFF-state, the results of the combined quantum mechanics{molecular mechanics (QM/MM) calculations show that chromophore photoisomerization happens in its neutral form and is followed by ground state deprotonation. This is in agreement with a very recent ultrafast absorption spectroscopy study for IrisFP, and studies on Dronpa and other negative RSFPs. Although the experimental results in both proteins show the same steps, I found that they have diferent processes at the atomic level due to structural and electrostatical diferences, but leading to the same intermediates. This is in contrast to the ON-state where I get the same picture for the three proteins studied, identifying the conical intersection that quenches the uorescence and controls the photoswitching quantum yield. The major diference between the three proteins in terms of uorescence and photoswitching characteristics comes from the diferent sterical environment produced by the residue 159, which is a methionine in diferent isomers in IrisFP and Dronpa and a smaller threonine that allows a faster isomerization of the chromophore in Dronpa2.Rubio Secades, AngelWanko, Marius ConnyFísica de Materiales;;Materialen Fisika2017201720162016info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10810/20519reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoInglésinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-sa/4.0/(cc)2016 BRUNO TORCAL EMBEITA (cc by-sa 4.0)oai:addi.ehu.eus:10810/205192026-06-18T09:23:17Z
dc.title.none.fl_str_mv Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
title Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
spellingShingle Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
Torcal Embeita, Bruno
molecular spectroscopy
photochemistry
quantum theory
espectroscopia molecular
fotoquímica
teoría cuántica
title_short Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
title_full Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
title_fullStr Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
title_full_unstemmed Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
title_sort Theoretical study on the photoswitching mechanism of negative reversibly photoswitchable fluorescent proteins
dc.creator.none.fl_str_mv Torcal Embeita, Bruno
author Torcal Embeita, Bruno
author_facet Torcal Embeita, Bruno
author_role author
dc.contributor.none.fl_str_mv Rubio Secades, Angel
Wanko, Marius Conny
Física de Materiales;;Materialen Fisika
dc.subject.none.fl_str_mv molecular spectroscopy
photochemistry
quantum theory
espectroscopia molecular
fotoquímica
teoría cuántica
topic molecular spectroscopy
photochemistry
quantum theory
espectroscopia molecular
fotoquímica
teoría cuántica
description 105 p.
publishDate 2016
dc.date.none.fl_str_mv 2016
2016
2017
2017
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/20519
url http://hdl.handle.net/10810/20519
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-sa/4.0/
(cc)2016 BRUNO TORCAL EMBEITA (cc by-sa 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-sa/4.0/
(cc)2016 BRUNO TORCAL EMBEITA (cc by-sa 4.0)
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301603

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