Bioluminescence inhibition assay for the detection of hydroxylated polychlorinated biphenyls.
Hydroxylated polychlorinated biphenyls (OH-PCBs) are an important class of contaminants that mainly originate from polychlorinated biphenyl metabolism. They may conceivably be as dangerous and persistent as the parent compounds; most prominently, OH-PCBs are endocrine disruptors. Due to increasing evidence of the presence of OH-PCBs in the environment and in living organisms, including humans, and of their toxicity, methods of detection for OH-PCBs are needed in the environmental and medical fields. Herein, we describe the development and optimization of a protein-based inhibition assay for the quantification of OH-PCBs. Specifically, the photoprotein aequorin was utilized for the detection of OH-PCBs. We hypothesized that OH-PCBs interact with aequorin, and we established that OH-PCBs actually inhibit the bioluminescence of aequorin in a dose-dependent manner. We took advantage of this phenomenon to develop an assay that is capable of detecting a wide variety of OH-PCBs with a range of detection limits, the best detection limit being 11 nM for the compound 2-hydroxy-2',3,4',5',6-pentachorobiphenyl. The viability of this system for the screening of OH-PCBs in spiked biological and environmental samples was also established. We envision the implementation of this novel bioluminescence inhibition assay as a rapid, sensitive, and cost-effective method for monitoring OH-PCBs. Furthermore, to the best of our knowledge, this is the first time aequorin has been employed to detect an analyte by the inhibition of its bioluminescence reaction. Hence, this strategy may prove to be a general approach for the development of a new generation of protein-based inhibition assays.
Department of Chemistry, University of Kentucky, Lexington, 40506-0055, United States.
Krystal
K
Teasley Hamorsky
C Mark
CM
Ensor
Emre
E
Dikici
Patrizia
P
Pasini
Leonidas
L
Bachas
Sylvia
S
Daunert
D000331:Aequorin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D052244:Endocrine Disruptors / Q000032:analysis / Q000378:metabolism; D004785:Environmental Pollutants / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D007093:Imidazoles / Q000737:chemistry; D008163:Luminescent Measurements; D011078:Polychlorinated Biphenyls / Q000032:analysis* / Q000378:metabolism; D011719:Pyrazines / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D052244:Endocrine Disruptors; D004785:Environmental Pollutants; D007093:Imidazoles; D011719:Pyrazines; D011994:Recombinant Proteins; C017144:coelenterazine; D000331:Aequorin; D011078:Polychlorinated Biphenyls; D004121:Dimethyl Sulfoxide