Effects of Currently Used Antifouling Booster Biocides on Green Fluorescent Proteins in Anemonia Viridis

The Mediterranean ◽

Green Fluorescent ◽

Anemonia Viridis

Abstract Some of the antifouling booster biocides affects the marine ecosystem negatively. The booster biocides which are resistant to degradation are accumulated in the sediment of the oceans. One of the sedentary organisms in the Mediterranean Sea is Anemonia viridis. The aim of this study is to show the toxicities of common biocides such as irgarol, seanine-211, zinc omadine, and acticide on the fluorescence by GFPs of A.viridis. The decreases in the fluorescence intensities of the GFP were measured within different booster biocide concentrations. The results show that fluorescent intensities of GFP proteins decreased more than 50 percent when they are exposed to different concentrations of irgarol, zinc omadine, acticide. In conclusion, ecosystem health should be prioritized when new antifouling paint compositions are proposed. From the results, it seems that A.viridis can be considered as a vulnerable organism and also it is sensitive to booster biocides within self-polishing antifouling paint formulations.

Functional expression and FRET analysis of green fluorescent proteins fused to G-protein subunits in rat sympathetic neurons

The Journal of Physiology ◽

10.1113/jphysiol.2001.013107 ◽

2001 ◽

Vol 537 (3) ◽

pp. 679-692 ◽

Cited By ~ 36

Author(s):

V. Ruiz-Velasco ◽

S. R Ikeda

Keyword(s):

G Protein ◽

Fluorescent Proteins ◽

Sympathetic Neurons ◽

Functional Expression ◽

Protein Subunits ◽

Coherent Dynamics of Photoexcited Green Fluorescent Proteins

Physical Review Letters ◽

10.1103/physrevlett.86.3439 ◽

2001 ◽

Vol 86 (15) ◽

pp. 3439-3442 ◽

Cited By ~ 38

Author(s):

Riccardo A. G. Cinelli ◽

Valentina Tozzini ◽

Vittorio Pellegrini ◽

Fabio Beltram ◽

Giulio Cerullo ◽

...

Keyword(s):

Fluorescent Proteins ◽

Coherent Dynamics ◽

Shedding light on the dark and weakly fluorescent states of green fluorescent proteins

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.96.11.6177 ◽

1999 ◽

Vol 96 (11) ◽

pp. 6177-6182 ◽

Cited By ~ 248

Author(s):

W. Weber ◽

V. Helms ◽

J. A. McCammon ◽

P. W. Langhoff

Keyword(s):

Fluorescent Proteins ◽

Mechanism and bottlenecks in strand photodissociation of split green fluorescent proteins (GFPs)

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1618087114 ◽

2017 ◽

Vol 114 (11) ◽

pp. E2146-E2155 ◽

Cited By ~ 7

Author(s):

Chi-Yun Lin ◽

Johan Both ◽

Keunbong Do ◽

Steven G. Boxer

Keyword(s):

Quantum Yield ◽

Protein Interactions ◽

Fluorescent Proteins ◽

Point Mutations ◽

Photoactive Yellow Protein ◽

Protein Protein Interactions ◽

Low Efficiency ◽

Green Fluorescent ◽

Rate Limiting

Split GFPs have been widely applied for monitoring protein–protein interactions by expressing GFPs as two or more constituent parts linked to separate proteins that only fluoresce on complementing with one another. Although this complementation is typically irreversible, it has been shown previously that light accelerates dissociation of a noncovalently attached β-strand from a circularly permuted split GFP, allowing the interaction to be reversible. Reversible complementation is desirable, but photodissociation has too low of an efficiency (quantum yield <1%) to be useful as an optogenetic tool. Understanding the physical origins of this low efficiency can provide strategies to improve it. We elucidated the mechanism of strand photodissociation by measuring the dependence of its rate on light intensity and point mutations. The results show that strand photodissociation is a two-step process involving light-activated cis-trans isomerization of the chromophore followed by light-independent strand dissociation. The dependence of the rate on temperature was then used to establish a potential energy surface (PES) diagram along the photodissociation reaction coordinate. The resulting energetics–function model reveals the rate-limiting process to be the transition from the electronic excited-state to the ground-state PES accompanying cis-trans isomerization. Comparisons between split GFPs and other photosensory proteins, like photoactive yellow protein and rhodopsin, provide potential strategies for improving the photodissociation quantum yield.

Multiphoton Bleaching of Red Fluorescent Proteins and the Ways to Reduce It

International Journal of Molecular Sciences ◽

10.3390/ijms23020770 ◽

2022 ◽

Vol 23 (2) ◽

pp. 770

Author(s):

Mikhail Drobizhev ◽

Rosana S. Molina ◽

Jacob Franklin

Keyword(s):

Chemical Structure ◽

Fluorescent Proteins ◽

Multiphoton Ionization ◽

Excitation Power ◽

Photon Absorption ◽

Two Photon Absorption ◽

Two Photon ◽

Red Fluorescent Proteins ◽

Red fluorescent proteins and biosensors built upon them are potentially beneficial for two-photon laser microscopy (TPLM) because they can image deeper layers of tissue, compared to green fluorescent proteins. However, some publications report on their very fast photobleaching, especially upon excitation at 750–800 nm. Here we study the multiphoton bleaching properties of mCherry, mPlum, tdTomato, and jREX-GECO1, measuring power dependences of photobleaching rates K at different excitation wavelengths across the whole two-photon absorption spectrum. Although all these proteins contain the chromophore with the same chemical structure, the mechanisms of their multiphoton bleaching are different. The number of photons required to initiate a photochemical reaction varies, depending on wavelength and power, from 2 (all four proteins) to 3 (jREX-GECO1) to 4 (mCherry, mPlum, tdTomato), and even up to 8 (tdTomato). We found that at sufficiently low excitation power P, the rate K often follows a quadratic power dependence, that turns into higher order dependence (K~Pα with α > 2) when the power surpasses a particular threshold P*. An optimum intensity for TPLM is close to the P*, because it provides the highest signal-to-background ratio and any further reduction of laser intensity would not improve the fluorescence/bleaching rate ratio. Additionally, one should avoid using wavelengths shorter than a particular threshold to avoid fast bleaching due to multiphoton ionization.