Ultramarine, a Chromoprotein Acceptor for Förster Resonance Energy Transfer

Pettikiriarachchi, Anne; Gong, Lan; Perugini, Matthew A.; Devenish, Rodney J.; Prescott, Mark

doi:10.1371/journal.pone.0041028

Cited by 33 publications

(44 citation statements)

References 26 publications

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“…The dark yellow REACh [15] and intense blue Ultramarine [17] are reported to be suitable acceptors for the emission of EGFP. However, use of such dark acceptors to monitor FRET requires donor fluorescence emission lifetimes to be determined using sophisticated and expensive instrumentation.…”

Section: Discussionmentioning

confidence: 99%

“…The complexity of such multi-parameter experiments is limited by the number of different FPs whose emission can be separately detected. Although the availability of non-fluorescent genetically encoded acceptors such as REACh [15], [16] or Ultramarine [17] has the potential to increase the number of separate events that might be monitored in the same experiment, access to expensive instrumentation is required to determine fluorescence lifetimes and FRET.…”

Section: Introductionmentioning

confidence: 99%

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Phanta: A Non-Fluorescent Photochromic Acceptor for pcFRET

et al. 2013

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We have developed an orange non-fluorescent photochromic protein (quantum yield, 0.003) we call Phanta that is useful as an acceptor in pcFRET applications. Phanta can be repeatedly inter-converted between the two absorbing states by alternate exposure to cyan and violet light. The absorption spectra of Phanta in one absorbing state shows excellent overlap with the emission spectra of a number of donor green fluorescent proteins including the commonly used EGFP. We show that the Phanta-EGFP FRET pair is suitable for monitoring the activation of caspase 3 in live cells using readily available instrumentation and a simple protocol that requires the acquisition of two donor emission images corresponding to Phanta in each of its photoswitched states. This the first report of a genetically encoded non-fluorescent acceptor for pcFRET.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Phanta: A Non-Fluorescent Photochromic Acceptor for pcFRET

et al. 2013

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“…These results indicated that the FRET from mRuby2 to dark mCherry mutants was taking place. Furthermore, we paired Ultramarine, which is a known dark fluorescent protein12, with mRuby2, and found that FRET also occurs in the mRuby2-Ultramarine pair (Fig. 2c,d).…”

Section: Resultsmentioning

confidence: 96%

“…Notably, because such pairs require only a narrow range of wavelengths (550–650 nm), they facilitate dual observation with a green fluorescent dye or protein using the 500–550 nm range of wavelengths (Figs 3 and 4). Although chromoprotein named Ultramarine was reported as an acceptor for FLIM-FRET, and its spectral properties are similar to those of dark mCherrys12, the reduced cell-to-cell variability of dark mCherrys represents a big advantage over Ultramarine (Fig. 2e).…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, a dark green fluorescent protein called ShadowG has been developed and applied to dual observation of Ras FRET biosensor activity and mCherry-ERK translocation11. In addition to dark green/yellow fluorescent proteins, a red-shifted nonfluorescent protein named Ultramarine has been developed12. This protein has a broad red-shifted absorption spectrum (as compared to REACh) and an extremely low quantum yield (~0.001), potentially applicable to a wide range of fluorescent proteins for FRET measurement.…”

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confidence: 99%

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Dual observation of the ATP-evoked small GTPase activation and Ca2+ transient in astrocytes using a dark red fluorescent protein

Nakahata

Nabekura

Murakoshi

2016

Sci Rep

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Intracellular signal transduction involves a number of biochemical reactions, which largely consist of protein-protein interactions and protein conformational changes. Monitoring Förster resonance energy transfer (FRET) by fluorescence lifetime imaging microscopy (FLIM), called FLIM-FRET, is one of the best ways to visualize such protein dynamics. Here, we attempted to apply dark red fluorescent proteins with significantly smaller quantum yields. Application of the dark mCherry mutants to single-molecule FRET sensors revealed that these dark mCherry mutants are a good acceptor in a pair with mRuby2. Because the FRET measurement between mRuby2 and dark mCherry requires only the red region of wavelengths, it facilitates dual observation with other signaling sensors such as genetically encoded Ca2+ sensors. Taking advantage of this approach, we attempted dual observation of Ca2+ and Rho GTPase (RhoA and Cdc42) activities in astrocytes and found that ATP triggers both RhoA and Cdc42 activation. In early phase, while Cdc42 activity is independent of Ca2+ transient evoked by ATP, RhoA activity is Ca2+ dependent. Moreover, the transient Ca2+ upregulation triggers long-lasting Cdc42 and RhoA activities, thereby converting short-term Ca2+ signaling to long-term signaling. Thus, the new FRET pair should be useful for dual observation of intracellular biochemical reactions.

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Chromogenic fusion proteins as alternative textiles dyes

Watkins,

Moffitt,

Speight

et al. 2024

Biotech & Bioengineering

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The widespread adoption of fast fashion has led to a significant waste problem associated with discarded textiles. Using proteins to color textiles can serve as a sustainable alternative to chemical dyes as well as reduce the demand for new raw materials. Here, we explore the use of chromogenic fusion proteins, consisting of a chromoprotein and a carbohydrate‐binding module (CBM), as coloring agents for cellulose‐based textiles such as cotton. We examined the color properties of chromoproteins AeBlue, SpisPink and Ultramarine alone and fused to CBM under various conditions. AeBlue, SpisPink and Ultramarine exhibited visible color between pH 4‐9 and temperatures ranging from 4 to 45℃. Fusing CBM Clos from Clostridium thermocellum and CBM Ch2 from Trichoderma reesei to the chromoproteins had no effect on the chromoprotein color properties. Furthermore, binding assays showed that chromoprotein fusions did not affect binding of CBMs to cellulosic materials. Cotton samples bound with Ultramarine‐Clos exhibited visible purple color that faded progressively over time as the samples dried. Applying 10% 8000 polyethylene glycol to cotton samples markedly preserved the color over extended periods. Overall, this work highlights the potential of chromoprotein‐CBM fusions for textile dying which could be applied as a color maintenance technology or for reversible coloring of textiles for events or work wear, contributing to sustainable practices and introducing new creative opportunities for the industry.

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Ultramarine, a Chromoprotein Acceptor for Förster Resonance Energy Transfer

Cited by 33 publications

References 26 publications

Phanta: A Non-Fluorescent Photochromic Acceptor for pcFRET

Phanta: A Non-Fluorescent Photochromic Acceptor for pcFRET

Dual observation of the ATP-evoked small GTPase activation and Ca2+ transient in astrocytes using a dark red fluorescent protein

Chromogenic fusion proteins as alternative textiles dyes

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