Tryptophan-based Chromophore in Fluorescent Proteins Can Be Anionic

Overview

Journal Sci Rep

Specialty Science

Date 2012 Aug 31

PMID 22934131

Citations 11

Authors

Karen S Sarkisyan

Ilia V Yampolsky

Kyril M Solntsev

Sergey A Lukyanov

Konstantin A Lukyanov

Alexander S Mishin

Affiliations

Soon will be listed here.

Abstract

Cyan fluorescent proteins (CFP) with tryptophan66-based chromophore are widely used for live cell imaging. In contrast to green and red fluorescent proteins, no charged states of the CFP chromophore have been described. Here, we studied synthetic CFP chromophore and found that its indole group can be deprotonated rather easily (pKa 12.4).We then reproduced this effect in the CFP mCerulean by placing basic amino acids in the chromophore microenvironment. As a result, green-emitting variant with an anionic chromophore and key substitution Val61Lys was obtained. This is the first evidence strongly suggesting that tryptophan-based chromophores in fluorescent proteins can exist in an anionic charged state. Switching between protonated and deprotonated Trp66 in fluorescent proteins represents a new unexplored way to control their spectral properties.

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