Anthozoa red fluorescent protein in biosensing

Abstract

The identification and cloning of a red fluorescent protein (DsRed) obtained from Anthozoa corals has provided an alternative to commonly used green fluorescent proteins (GFPs) in bioanalytical and biomedical research. DsRed in tandem with GFPs has enhanced the feasibility of multicolor labeling studies. Properties of DsRed, for example high photostability, red-shifted fluorescence emission, and stability to pH changes have proven valuable in its use as a fluorescent tag in cell-biology applications. DsRed has some limitations, however. Its slow folding and tendency to form tetramers have been a hurdle. Several different mutational studies have been performed on DsRed to overcome these problems. In this paper, applications of DsRed in biosensing, specifically in FRET/BRET assays, whole-cell assays, and in biosensors, is discussed. In the future, construction of DsRed mutants with unique characteristics will further expand its applications in bioanalysis.

Mechanism of copper induced fluorescence quenching of red fluorescent protein, DsRed

“…These red fluorescent proteins form chromophores internally and share structural similarities with green fluorescent proteins (GFP) [1][2][3]. Isolation and characterization of these GFP-like red proteins has expanded the possible applications of fluorescent proteins into multi-color labeling, resonance energy transfer, and intracellular tracking studies [2,[4][5][6]. Like GFP, red fluorescent proteins have been mainly employed as genetically encoded fluorescent probes for cellular applications.…”

Molecular-Size Fluorescence Emitters

“…With molar absorption coefficient 57,000 M −1 cm −1 and quantum yield 79 % it competes with GFP and shows superior properties in terms of photostability and independence of spectra on pH in the range 5.0-12.0 (Shrestha and Deo 2006). Excited in the range 558-578 nm DsRed and its mutants display fluorescence with the band maxima ranging between 583 and 605 nm.…”

Computed tomography screening for lung cancer: Review of screening principles and update on current status

“…The discovery of the green fluorescent protein (GFP), from the jellyfish Aequorea victoria in 1962 1, has revolutionized the use of fluorescence imaging in the field of cellular and molecular biology 2. The chromophore, which is largely responsible for the optical properties of GFP and GFP‐like proteins, is formed by autocatalytic maturation of a tripeptide sequence within the protein 3, 4. Thus, the information needed for the synthesis of the chromophore resides in the corresponding gene, making these proteins well‐suited as genetically fused, fluorescent biomarkers.…”

“…The development of fluorescent protein (FP) mutants has made a range of fluorescent colors available 3 and these proteins have proved to be useful in real‐time studies of processes in living cells as reflected in their numerous applications. A few examples are monitoring of gene expression, simultaneous imaging of the cellular location and dynamics of several biomolecules as well as fluorescence‐based techniques, e.g., fluorescent resonance energy‐transfer.…”

“…A few examples are monitoring of gene expression, simultaneous imaging of the cellular location and dynamics of several biomolecules as well as fluorescence‐based techniques, e.g., fluorescent resonance energy‐transfer. These procedures have provided important information about the concerted action of biomolecules, contributing to the elucidation of their biological function 2, 3, 5, 6. Despite the extensive use of the aforementioned methodologies, there are still challenges to be solved, e.g., increasing the brightness of the FPs, color‐tuning of the emission to the near IR‐region 3, and enhancing the two‐photon cross sections.…”