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Correlation between fluorescence and structure in the orange-emitting GFP-like protein, monomeric Kusabira Orange

Paper ID Volume ID Publish Year Pages File Format Full-Text
30301 44468 2014 7 PDF Available
Title
Correlation between fluorescence and structure in the orange-emitting GFP-like protein, monomeric Kusabira Orange
Abstract

•We described the thermal denaturation process of mKO.•mKO shows a high thermostability and an irreversible three-state unfolding process.•Changes in chromophore protonation lead to variations of fluorescence intensity.•Changes in chromophore protonation are coupled to small structural rearrangements.

The mKO is the monomeric version of Kusabira Orange, a GFP-like protein emitting bright orange fluorescence at 559 nm. This protein shows the characteristic β-barrel motif typical of the fluorescent protein family which it belongs to, similar spectral properties to the tetrameric form and an exceptional photo-stability to pH changes. Here, we demonstrate that mKO in solution at physiological pH exhibits a secondary structure analogue to that of the crystal. Moreover, we describe the thermal unfolding, revealing an outstanding structural stability with a denaturation temperature close to 90 °C and identifying the existence of a thermodynamic intermediate. The denaturation process of mKO results to be absolutely irreversible because of the complete lost of the native structure and the consequent aggregation, while the presence of the intermediate state is most likely due to coexistence of two different species of mKO, with protonated and deprotonated chromophore respectively, that affects the fluorescence properties and the structural stability of the protein.

Keywords
mKO; Fluorescent proteins; Thermo-stability; Protein structure; FTIR
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Correlation between fluorescence and structure in the orange-emitting GFP-like protein, monomeric Kusabira Orange
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology B: Biology - Volume 138, 5 September 2014, Pages 223–229
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us