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Exploring chromophore-protein interactions in fluorescent protein cmFP512 from Cerianthus membranaceus: X-ray structure analysis and optical spectroscopy

Nienhaus, K., Renzi, F., Vallone, B., Wiedenmann, J. and Nienhaus, G.U. (2006) Exploring chromophore-protein interactions in fluorescent protein cmFP512 from Cerianthus membranaceus: X-ray structure analysis and optical spectroscopy Biochemistry, 45, (43), pp. 12942-12953. (doi:10.1021/bi060885c S0006-2960(06)00885-3).

Record type: Article

Abstract

Autofluorescent proteins of the GFP family all share the same three-dimensional -can fold; yet they exhibit widely different optical properties, arising either from chemical modification of the chromophore itself or from specific interactions of the chromophore with the surrounding protein moiety. Here we present a structural and spectroscopic characterization of the green fluorescent protein cmFP512 from Cerianthus membranaceus, a nonbioluminescent, azooxanthellate cnidarian, which has only ~22% sequence identity with Aequorea victoria GFP. The X-ray structure, obtained by molecular replacement at a resolution of 1. 35 Å, shows the chromophore, formed from the tripeptide Gln-Tyr-Gly, in a hydrogen-bonded cage in the center of an 11-stranded -barrel, tightly restrained by adjacent residues and structural water molecules. It exists in a neutral (A) and an anionic (B) species, with absorption/emission maxima at 392/460 (pH 5) and 503/512 nm (pH 7). Their fractional populations and peak positions depend sensitively on pH, reflecting protonation of groups adjacent to the chromophore. The pH dependence of the spectra is explained by a protonation mechanism involving a hydrogen-bonded cluster of charged/polar groups. Cryospectroscopy at 12 K was also performed to analyze the vibronic coupling of the electronic transitions.

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Published date: 10 October 2006

Identifiers

Local EPrints ID: 51175
URI: http://eprints.soton.ac.uk/id/eprint/51175
ISSN: 0006-2960
PURE UUID: 17af0ea6-06cd-4dd7-825c-8efeb47ec02a
ORCID for J. Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

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Date deposited: 08 May 2008
Last modified: 11 Jul 2017 09:46

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Contributors

Author: K. Nienhaus
Author: F. Renzi
Author: B. Vallone
Author: J. Wiedenmann ORCID iD
Author: G.U. Nienhaus

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