Bioluminescent and Fluorescent Proteins: Molecular Mechanisms and Modern Applications (original) (raw)

The Growing and Glowing Toolbox of Fluorescent and Photoactive Proteins

Trends in biochemical sciences, 2016

Over the past 20 years, protein engineering has been extensively used to improve and modify the fundamental properties of fluorescent proteins (FPs) with the goal of adapting them for a fantastic range of applications. FPs have been modified by a combination of rational design, structure-based mutagenesis, and countless cycles of directed evolution (gene diversification followed by selection of clones with desired properties) that have collectively pushed the properties to photophysical and biochemical extremes. In this review, we provide both a summary of the progress that has been made during the past two decades, and a broad overview of the current state of FP development and applications in mammalian systems.

Engineering Bioluminescent Proteins: Expanding their Analytical Potential

Analytical Chemistry, 2009

Bioluminescence has been observed in nature since the dawn of time, but now, scientists are harnessing it for analytical applications. Laura Rowe, Emre Dikici, and Sylvia Daunert of the University of Kentucky describe the origins of bioluminescent proteins and explore their uses in the modern chemistry laboratory. The cover features spectra of bioluminescent light superimposed on an image of jellyfish, which are a common source of bioluminescent proteins. Images courtesy of Emre Dikici and Shutterstock.

Protein-protein complexation in bioluminescence

Protein & Cell, 2011

In this review we summarize the progress made towards understanding the role of protein-protein interactions in the function of various bioluminescence systems of marine organisms, including bacteria, jellyfish and soft corals, with particular focus on methodology used to detect and characterize these interactions. In some bioluminescence systems, protein-protein interactions involve an "accessory protein" whereby a stored substrate is efficiently delivered to the bioluminescent enzyme luciferase. Other types of complexation mediate energy transfer to an "antenna protein" altering the color and quantum yield of a bioluminescence reaction. Spatial structures of the complexes reveal an important role of electrostatic forces in governing the corresponding weak interactions and define the nature of the interaction surfaces. The most reliable structural model is available for the protein-protein complex of the Ca 2+-regulated photoprotein clytin and green-fluorescent protein (GFP) from the jellyfish Clytia gregaria, solved by means of Xray crystallography, NMR mapping and molecular docking. This provides an example of the potential strategies in studying the transient complexes involved in bioluminescence. It is emphasized that structural studies such as these can provide valuable insight into the detailed mechanism of bioluminescence.

A never ending race for new and improved fluorescent proteins

BMC Biology, 2012

Fluorescence and bioluminescence occur in terrestrial and aquatic organisms, but are especially common in the marine environment, where they serve diverse functions such as detecting or luring prey in dark environments, the attraction of mates, or a means to evade predation . Proteins are key to these phenomena and the first autofluorescent proteins were identified in the crystal jelly Aequorea victoria . The subsequent impact on bio logical research of these fluorescent proteins (FPs), with A. victoria green FP (avGFP) as the founding member, cannot be overstated. The great importance of FPs in biological research was recognized with the Nobel Prize in Chemistry to Osamu Shimomura, Martin Chalfie, and Roger Y Tsien in 2008 [4]. In a study just published in BMC Biotechnology, Mann et al. compare orange FP variants that were expressed transiently and stably in plants, raising the issue of what questions need to be asked in the selection of fluorophores. Here we sum marize the key questions that need to be addressed to find an optimal fluorophore for a given application in plant biology and beyond.

Directionality of light absorption and emission in representative fluorescent proteins

2020

Significance Fluorescent proteins have been used extensively in many areas of life sciences. Many of their applications rely on their various biophysical properties, such as excitation and emission wavelengths, excited state lifetimes, or sensitivity to the molecular environment. One aspect of fluorescent proteins that has remained largely neglected is the directionality of their optical properties. In the present work, we describe our experimental determination of the directionality of light absorption and emission in several commonly used fluorescent proteins. Our findings improve our understanding of fundamental properties of fluorescent proteins, and expand the possibilities of development and applications of genetically encoded fluorescent probes.

Advances in engineering of fluorescent proteins and photoactivatable proteins with red emission

Current Opinion in Chemical Biology, 2010

Monomeric fluorescent proteins of different colors are widely used to study behavior and targeting of proteins in living cells. Fluorescent proteins that irreversibly change their spectral properties in response to light irradiation of a specific wavelength, or photoactivate, have become increasingly popular to image intracellular dynamics and super-resolution protein localization. Until recently, however, no optimized monomeric red fluorescent proteins and red photoactivatable proteins have been available. Furthermore, monomeric fluorescent proteins, which change emission from blue to red simply with time, so-called fluorescent timers, were developed to study protein age and turnover. Understanding of chemical mechanisms of the chromophore maturation or photoactivation into a red form will further advance engineering of fluorescent timers and photoactivatable proteins with enhanced and novel properties.