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A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein

Citation

Rodriguez, EA and Tran, GN and Gross, LA and Crisp, JL and Shu, X and Lin, JY and Tsien, RY, A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein, Nature Methods: techniques for life scientists and chemists, 13, (9) pp. 763-769. ISSN 1548-7091 (2016) [Refereed Article]

Copyright Statement

Copyright 2016 Nature America, Inc.

DOI: doi:10.1038/nmeth.3935

Abstract

Far-red fluorescent proteins (FPs) are desirable for in vivo imaging because with these molecules less light is scattered, absorbed, or re-emitted by endogenous biomolecules compared with cyan, green, yellow, and orange FPs. We developed a new class of FP from an allophycocyanin α-subunit (APCα). Native APC requires a lyase to incorporate phycocyanobilin. The evolved FP, which we named small ultra-red FP (smURFP), covalently attaches a biliverdin (BV) chromophore without a lyase, and has 642/670-nm excitation–emission peaks, a large extinction coefficient (180,000 M−1cm−1) and quantum yield (18%), and photostability comparable to that of eGFP. smURFP has significantly greater BV incorporation rate and protein stability than the bacteriophytochrome (BPH) FPs. Moreover, BV supply is limited by membrane permeability, and smURFPs (but not BPH FPs) can incorporate a more membrane-permeant BV analog, making smURFP fluorescence comparable to that of FPs from jellyfish or coral. A far-red and near-infrared fluorescent cell cycle indicator was created with smURFP and a BPH FP.

Item Details

Item Type:Refereed Article
Keywords:fluorescent protein, microscopy, protein engineering and design
Research Division:Technology
Research Group:Medical Biotechnology
Research Field:Medical Molecular Engineering of Nucleic Acids and Proteins
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Biological Sciences
Author:Lin, JY (Dr John Lin)
ID Code:111383
Year Published:2016
Web of Science® Times Cited:21
Deposited By:Medicine (Discipline)
Deposited On:2016-09-09
Last Modified:2017-11-07
Downloads:0

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