Bioluminescent Assay of Farnesyl Protein Transferase (FPTase) Using Bacterial Luciferase from Vibrio harveyi

Abstract

Instead of the usual assay method of FPTase using radioisotope labelled farnesyl pyrophosphate and synthetic peptide (-CXXA-) as substrates, a luminous assay method using bacterial luciferase as substrate was designed. Luciferase from marine luminous bacterium V. harveyi has highly active Cys. residue in its active site and lose activity by any modifying agent such as 2-bromo-decanal. In the presence of FPTase, added farnesyl pyrophosphate acts as farnesylation agent to bacterial luciferase Cys-106 sulfhydryl group and inactivate the luciferase activity completely while other luciferase from V. fischeri without Cys-106 sulfhydryl group didn't show irreversible inhibition. This result can be developed to a new non-radioisotopic assay method of FPTase which is a very important target for anticancer drug development