Bioluminescence assay for detecting cell surface membrane protein expression.
We have developed a method to measure the amounts of cell surface-expressed membrane proteins with bioluminescence. Dinoflagellate luciferase was expressed on the surface of a mammalian cell as a chimeric fusion protein with a membrane protein of interest. Using a membrane-impermeable substrate to quantify the membrane-displayed luciferase, the expression of the membrane protein on the cell surface was determined. By inclusion of a quenching step for the luminescent activity of luciferase on the cell surface, we were able to monitor the membrane protein expression kinetics by measuring the luminescence recovery from the cell surface after quenching. The reported methods provide a convenient way to monitor the kinetics of expression and transport of membrane proteins to the cell surface. It is applicable to the high-throughput analysis of drugs or drug candidates concerning their effects on membrane protein expression.
Kato
Mieko
M
Signaling Molecules Research Group, Neuroscience Research Institute, National Institutes of Advanced Industrial Science and Technology, Tsukuba, 1-1-1 Higashi, Tsukuba, Japan.
Chiba
Tomoki
T
Li
Min
M
Hanyu
Yoshiro
Y
D001681:Biological Assay / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008156:Luciferases / Q000032:analysis*; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*
D008164:Luminescent Proteins; D008565:Membrane Proteins; D008156:Luciferases