Circular dichroism spectroscopy of fluorescent proteins.

Abstract:

:Circular dichroism (CD) spectra have been obtained from several variants of green fluorescent protein: blue fluorescent protein (BFP), enhanced cyan fluorescent protein (CFP), enhanced green fluorescent protein (GFP), enhanced yellow fluorescent protein (YFP), all from Aequorea victoria, and the red fluorescent protein from the coral species Discosoma (DsRed). We demonstrate that CD spectra in the spectral fingerprint region of the chromophore yield spectra that after normalization are not coincident with the normalized absorbance spectra of GFP, YFP and DsRed. On the other hand, the CD spectra of BFP and CFP coincide with the absorbance spectra. The resolution of absorption and CD spectra into Gaussian bands confirmed the location of the different electronic band positions of GFP and YFP as reported in the literature using other techniques. In the case of BFP and CFP the location of Gaussian bands provided information of the vibrational progression of the electronic absorption bands. The CD spectrum of DsRed is anomalous in the sense that the major CD band has a clear excitonic character. Far-UV CD spectra of GFP confirmed the presence of the high beta-sheet content of the polypeptide chain in the three-dimensional structure.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Visser NV,Hink MA,Borst JW,van der Krogt GN,Visser AJ

doi

10.1016/s0014-5793(02)02808-9

keywords:

subject

Has Abstract

pub_date

2002-06-19 00:00:00

pages

31-5

issue

1-3

eissn

0014-5793

issn

1873-3468

pii

S0014579302028089

journal_volume

521

pub_type

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