Bioluminescence Assay for Detecting Cell Surface Membrane Protein Expression

Abstract

We have developed a method to measure the amounts of cell surfaceexpressed 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.

Applications of bioluminescence in biotechnology and beyond

“…B135 kDa 129 131 measuring quantities of cell-surface expressed membrane proteins, 132 and pregnancy-specific glycoproteins in HeLa cells to investigate cell senescence. 133 Quantum yield not determined.…”

Microalgae: Current Research and Applications

“…Single domains of dinoflagellate LCF fused to genes of interest in mammalian cells produce fusion protein whose abundance can be monitored by a rapid bioluminescence assay, thus allowing for monitoring gene expression levels in the tagged genes (Suzuki et al, 2005). In a subsequent study, Kato et al (2011) used dinoflagellate LCF to monitor the expression and kinetics of a cell membrane protein. The authors fused LCF to the extracellular part of platelet-derived growth factor receptor transmembrane protein of a mammalian cell, which could then be detected by the addition of luciferin.…”

Inhibition of Gaussia luciferase Activity by a Monoclonal Antibody

“…Many types of luciferases, such as aequorin from jellyfish and luciferases from the firefly, crick beetle, Renilla and Gaussia princeps, are used [4][5][6][7]. A high-throughput assay for membrane protein expression on the cell surface has been developed with dinoflagellate luciferase [10]. A high-throughput assay for membrane protein expression on the cell surface has been developed with dinoflagellate luciferase [10].…”