The photophysical mechanism underlying the photochromic behavior of green fluorescent protein (GFP) mutants is investigated by means of preresonant Raman spectroscopy and model calculations. The studied molecules are reversibly switchable fluorophores that can be repeatedly converted between fluorescent and nonfluorescent states by irradiation and are attracting a broad interest for a number of new applications. Experimental results on chemically synthesized isolated chromophores are analyzed within a density functional theory approach that allows us to link the detected vibrational modes to specific ground-state configurations before and after photoconversion. These data are compared to results obtained for the case of complete folded proteins including the same chromophores. Our results highlight the impact of chromophore cis-trans isomerization and protonation state in the photophysics of these proteins and provide useful guidelines for novel mutant design.

Raman study of chromophore states in photochromic fluorescent proteins

Luin, Stefano
;
Beltram, Fabio
2009

Abstract

The photophysical mechanism underlying the photochromic behavior of green fluorescent protein (GFP) mutants is investigated by means of preresonant Raman spectroscopy and model calculations. The studied molecules are reversibly switchable fluorophores that can be repeatedly converted between fluorescent and nonfluorescent states by irradiation and are attracting a broad interest for a number of new applications. Experimental results on chemically synthesized isolated chromophores are analyzed within a density functional theory approach that allows us to link the detected vibrational modes to specific ground-state configurations before and after photoconversion. These data are compared to results obtained for the case of complete folded proteins including the same chromophores. Our results highlight the impact of chromophore cis-trans isomerization and protonation state in the photophysics of these proteins and provide useful guidelines for novel mutant design.
2009
Settore FIS/03 - Fisica della Materia
Density-functional theory; vibrational properties; structural basis; light; decarboxylation; molecules; dark; green fluorescent proteins; photochemical processes; protein folding
File in questo prodotto:
File Dimensione Formato  
ja804504b_withSI.pdf

Accesso chiuso

Descrizione: Article + Supplementary Material
Tipologia: Published version
Licenza: Non pubblico
Dimensione 1.79 MB
Formato Adobe PDF
1.79 MB Adobe PDF   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11384/8189
Citazioni
  • ???jsp.display-item.citation.pmc??? 7
  • Scopus 35
  • ???jsp.display-item.citation.isi??? 35
  • OpenAlex ND
social impact