Applications of luciferin biosynthesis: Bioluminescence assays for l-cysteine and luciferase.
Based on the biosynthetic pathway of firefly bioluminescence substrate D-luciferin, the concentration of L-cysteine can be quantified using a simple protocol and a conventional luminescence detector. The lower limit of quantification (signal/noise ratio [S/N]=10) was 0.26 microM. Using our method, the total amount of free/reduced and disulfide/oxidized L-cysteine could be measured successfully in human serum. In addition, biosynthetic precursors such as 2-cyano-6-hydroxybenzothiazole and L-luciferin could replace d-luciferin in the cell-based luciferase assay. Our results suggest that the bioluminescence reaction associated with the biosynthesis of bioluminescence substrates can provide a fast and cost-effective assay method.
Niwa
Kazuki
K
National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8568, Japan.
Nakajima
Yoshihiro
Y
Ohmiya
Yoshihiro
Y
D052160:Benzothiazoles / Q000737:chemistry / Q000378:metabolism; D003545:Cysteine / Q000097:blood* / Q000378:metabolism; D049454:Firefly Luciferin / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D014176:Protein Biosynthesis
D052160:Benzothiazoles; D049454:Firefly Luciferin; D008156:Luciferases; C005465:benzothiazole; D003545:Cysteine