QUANTIFICATION OF MUTATED RAS:RAF COMPLEXES IN VIVO
Strategy to systematically quantify protein complex formation of cancer relevant, full length, and mutated RAS:RAF in vivo. The presented extendable reporter system facilitates the identification of lead molecules which reduce/prevent either directly or allosterically the formation of carcinogenic H-,K-,N-RAS : A-,B-,C-RAF complexes in vivo.
Deregulations of components of the RAS-RAF-ERK signaling pathway favor tumor formation. Recurrent oncogenic point mutations cause constitutive RAS or RAF activation. (i) Due to missing binding pockets for physical small molecules interactions mutant RAS has been termed ‘undrugable’. Proliferative RAS signaling involves the binary interaction with the RAF kinase. (ii) RAF activation depends on the interaction with GTP-bound RAS. BRAF is the most frequently mutated oncogene in the kinase superfamily. Perturbation of specific RAS:RAF complexes is one feasible strategy to interfere with oncogenic functions of mutated RAS and/or RAF.
We present a strategy to identify small molecules/peptides/pathways which affect mutated or wild type RAS:RAF complexes. We present an extendable cell based reporter system for RAS:RAF complexes to:
- Screen peptidic or small molecule inhibitors of deregulated RAS:RAF complexes
- Identify OFF‐target effects of lead molecules/approved drugs acting on RAF
- The three human Ras genes (HRas, KRas & NRas) are the most common oncogenes in human cancer
- About 50 % of melanomas harbor activating BRAF mutations (300 diffe rent RAF mutations have been identified and can be tested)
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StichworteCancer, RAS RAF, Reporter system, RAF kinase