KRAS and other Ras-family enzymes are an important component of over 30% of human cancers, however, no effective therapeutics targeting Ras or Ras-driven cancers are currently available. The production of Ras proteins in vitro is required for the identification and characterization of Ras targeting drugs. An important step in producing the Ras protein involves prenylation of the C-terminus of the protein via farnesyltransferase, a modification that does not occur in prokaryotic organisms. Previous attempts to generate properly processed Ras in eukaryotic cells has produced only low levels of protein which has not been useful for structural studies or biochemical work.
Researchers at the FNL Protein Expression Lab developed reagents that can be used to produce prenylated proteins such as KRAS in high yield. Available for licensing are baculovirus vectors for overexpression of human FNTA and FNTB; recombinant baculovirus genomes (bacmids) containing overexpression constructs of FNTA, FNTB, or combinations thereof; and DH10BAC cell lines that contain the modified bacmids and the baculovirus vectors. Baculovirus reagents for high-yield (>10 mg/L) production of properly processed KRAS are also available.
- Production of proteins for use in identification and characterization of drugs targeting Ras
- Potential to generate other prenylated proteins
- Production of prenylated proteins, such as Ras, in high yield
- Prenylated proteins produced using this method show membrane interaction activities that recapitulate in vivo activities not observed with bacterial produced proteins
Dominic Esposito (NCI), Carissa Grosse (NCI), William Gillette (NCI)
- Research Material: NIH will not pursue patent prosecution for this technology