Green Fluorescent Protein as a Biosensor for Toxic Compounds

Abstract

In this brief review, we present recent results in the development of fluorescencebased assays for the detection of compounds with cytotoxic, anticancer and antimicrobial properties. As other reviews have explored various aspects related to these topics, this review will focus on the use of the Green Fluorescent Protein (GFP) for the detection of potentially toxic and/or therapeutic compounds. Since high-throughput screening of chemical compounds can be both expensive and laborious, development of low cost and efficient cell-based assays to determine biological activity should greatly enhance the early screening process. In our recent studies, we have developed a couple of GFP-based assays for the rapid screening of compounds with cytotoxic and bacteriocidal properties. As will be described in more detail in subsequent sections, a new 96-well assay has recently been developed that allows for the simultaneous screening of test compounds on gram positive and negative bacteria as well as mammalian (human cancer) cells. Our results demonstrate that both mammalian cells and bacteria can be analyzed in tandem to rapidly determine which compounds are specifically toxic to one of these cell types. The parallel screening of both eukaryotic and prokaryotic cells was found to be feasible, reproducible, and cost effective. BRIEF OVERVIEW ON THE PROPERTIES OF GFPFew methods exist that allow the visualization of living cells as they undergo apoptotic or necrotic modes of death. Since labeling of cells with dyes is not always efficient and may lead to interference of cellular functions, the preferred method for non-invasive detection is via the use of fluorescent markers. A fluorescent marker that has achieved prominence in visualization assays is GFP and its derivatives. These

Do copper ions activate tyrosinase enzyme? A biosensor model for the solution

“…Biosensors could make monitoring and control in the biotech industry accurate and reproducible. Specific fields of interest are the bioprocess industry, the medical field, environmental detection of pollutants or toxins [14][15][16][17][18][19][20][21][22][23][24]. Some original and attractive biosensors were developed for the determination of copper such as microbial [25][26][27][28], carbon paste based [29][30][31][32] and optical [33,34].…”

Free energy calculations of protein-water complexes with Gromacs

“…We used Green fluorescent protein (GFP)-water complexes in our simulation. The Green fluorescent protein (GFP) [4][5][6][7] is an important proteins currently used as a marker of gene expression and protein localization, and as a bio-sensor [8]. The green fluorescent protein (GFP) is composed of 238 amino acid residues and a molecular weight of 26.9 kdalton that exhibits bright green fluorescence when exposed to light [9].…”

Designing a bacterial biosensor for detection of mercury in water solutions

“…Based on our knowledge, there are not any reports of building such a construct for biologically detecting mercury. There are whole-cell biosensors that use reporter systems and upstream mercury-inducible promoters (Ivask et al, 2001;Aguilera et al, 2006;Nagata et al, 2010). There are extensive reports of using bacterial biosensors since 1993; over time, the specificity and sensitivity of these biosensors have increased.…”