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Yeast screens for inhibitors of Ras-Raf interaction and characterization of MCP inhibitors of Ras-Raf interaction. 2006

Vladimir Khazak, and Juran Kato-Stankiewicz, and Fuyu Tamanoi, and Erica A Golemis
NexusPharma, Inc., Langhorne, Pennsylvania, USA.

Because of the central role of Ras in cancer cell signaling, there has been considerable interest in developing small molecule inhibitors of the Ras signaling pathways as potential chemotherapeutic agents. This chapter describes the use of a two-hybrid approach to identify the MCP compounds, small molecules that disrupt the interaction between Ras and its effector Raf. We first outline the reagent development and selection/counter selection methods required to successfully apply a two-hybrid approach to isolation of MCP compounds. Separately, we describe the collateral benefits of this screening approach in yielding novel antifungal compounds. We then discuss secondary physiological validation approaches to confirm the MCP compounds specifically target Ras-Raf signaling. Finally, we develop a decision tree for subsequent preclinical characterization and optimization of this class of pathway-targeted reagent.

UI MeSH Term Description Entries
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D047428 Protein Kinase Inhibitors Agents that inhibit PROTEIN KINASES. Protein Kinase Inhibitor,Inhibitor, Protein Kinase,Inhibitors, Protein Kinase,Kinase Inhibitor, Protein,Kinase Inhibitors, Protein
D048049 Extracellular Signal-Regulated MAP Kinases A mitogen-activated protein kinase subfamily that is widely expressed and plays a role in regulation of MEIOSIS; MITOSIS; and post mitotic functions in differentiated cells. The extracellular signal regulated MAP kinases are regulated by a broad variety of CELL SURFACE RECEPTORS and can be activated by certain CARCINOGENS. ERK MAP Kinase,ERK MAP Kinases,Extracellular Signal-Regulated Kinase,Extracellular Signal-Regulated Kinases,Extracellular Signal-Regulated MAP Kinase,MAP Kinases, Extracellular Signal-Regulated,Extracellular Signal Regulated Kinase,Extracellular Signal Regulated Kinases,Extracellular Signal Regulated MAP Kinase,Extracellular Signal Regulated MAP Kinases,Kinase, ERK MAP,Kinase, Extracellular Signal-Regulated,Kinases, Extracellular Signal-Regulated,MAP Kinase, ERK,MAP Kinases, Extracellular Signal Regulated,Signal-Regulated Kinase, Extracellular
D048490 raf Kinases A family of closely-related serine-threonine kinases that were originally identified as the cellular homologs of the retrovirus-derived V-RAF KINASES. They are MAP kinase kinase kinases that play important roles in SIGNAL TRANSDUCTION. MIL-RAF Proteins,mil-raf Protein Kinases,raf Kinase,raf MAP Kinase Kinase Kinases,Proto-Oncogene Proteins raf,raf Proto-Oncogene Proteins,raf Serine-Theonine Protein Kinases,Kinase, raf,Kinases, mil-raf Protein,Kinases, raf,MIL RAF Proteins,Protein Kinases, mil-raf,Proteins raf, Proto-Oncogene,Proto Oncogene Proteins raf,Proto-Oncogene Proteins, raf,mil raf Protein Kinases,raf Proto Oncogene Proteins,raf Serine Theonine Protein Kinases
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D018631 ras Proteins Small, monomeric GTP-binding proteins encoded by ras genes (GENES, RAS). The protooncogene-derived protein, PROTO-ONCOGENE PROTEIN P21(RAS), plays a role in normal cellular growth, differentiation and development. The oncogene-derived protein ONCOGENE PROTEIN P21(RAS) can play a role in aberrant cellular regulation during neoplastic cell transformation (CELL TRANSFORMATION, NEOPLASTIC). This enzyme was formerly listed as EC 3.6.1.47. Gene Products, ras,ras GTPase,ras Protein,ras GTPases,GTPase, ras,GTPases, ras,Protein, ras,ras Gene Products
D020798 Two-Hybrid System Techniques Screening techniques first developed in yeast to identify genes encoding interacting proteins. Variations are used to evaluate interplay between proteins and other molecules. Two-hybrid techniques refer to analysis for protein-protein interactions, one-hybrid for DNA-protein interactions, three-hybrid interactions for RNA-protein interactions or ligand-based interactions. Reverse n-hybrid techniques refer to analysis for mutations or other small molecules that dissociate known interactions. One-Hybrid System Techniques,Reverse One-Hybrid System Techniques,Reverse Two-Hybrid System Techniques,Three-Hybrid System Techniques,Yeast Two-Hybrid Assay,Yeast Two-Hybrid System Techniques,One-Hybrid System Technics,Reverse Three-Hybrid System Techniques,Three-Hybrid System Technics,Tri-Hybrid System Techniques,Two-Hybrid Assay,Two-Hybrid Method,Two-Hybrid System Technics,Yeast One-Hybrid System Techniques,Yeast Three-Hybrid Assay,Yeast Three-Hybrid System,Yeast Three-Hybrid System Techniques,Yeast Two-Hybrid System,n-Hybrid System Techniques,Assay, Two-Hybrid,Assay, Yeast Three-Hybrid,Assay, Yeast Two-Hybrid,Assays, Two-Hybrid,Assays, Yeast Three-Hybrid,Assays, Yeast Two-Hybrid,Method, Two-Hybrid,Methods, Two-Hybrid,One Hybrid System Technics,One Hybrid System Techniques,One-Hybrid System Technic,One-Hybrid System Technique,Reverse One Hybrid System Techniques,Reverse Three Hybrid System Techniques,Reverse Two Hybrid System Techniques,System Technique, n-Hybrid,System Techniques, n-Hybrid,System, Yeast Three-Hybrid,System, Yeast Two-Hybrid,Systems, Yeast Three-Hybrid,Systems, Yeast Two-Hybrid,Technic, One-Hybrid System,Technic, Three-Hybrid System,Technic, Two-Hybrid System,Technics, One-Hybrid System,Technics, Three-Hybrid System,Technics, Two-Hybrid System,Technique, One-Hybrid System,Technique, Three-Hybrid System,Technique, Tri-Hybrid System,Technique, Two-Hybrid System,Technique, n-Hybrid System,Techniques, One-Hybrid System,Techniques, Three-Hybrid System,Techniques, Tri-Hybrid System,Techniques, Two-Hybrid System,Techniques, n-Hybrid System,Three Hybrid System Technics,Three Hybrid System Techniques,Three-Hybrid Assay, Yeast,Three-Hybrid Assays, Yeast,Three-Hybrid System Technic,Three-Hybrid System Technique,Three-Hybrid System, Yeast,Three-Hybrid Systems, Yeast,Tri Hybrid System Techniques,Tri-Hybrid System Technique,Two Hybrid Assay,Two Hybrid Method,Two Hybrid System Technics,Two Hybrid System Techniques,Two-Hybrid Assay, Yeast,Two-Hybrid Assays,Two-Hybrid Assays, Yeast,Two-Hybrid Methods,Two-Hybrid System Technic,Two-Hybrid System Technique,Two-Hybrid System, Yeast,Two-Hybrid Systems, Yeast,Yeast One Hybrid System Techniques,Yeast Three Hybrid Assay,Yeast Three Hybrid System,Yeast Three Hybrid System Techniques,Yeast Three-Hybrid Assays,Yeast Three-Hybrid Systems,Yeast Two Hybrid Assay,Yeast Two Hybrid System,Yeast Two Hybrid System Techniques,Yeast Two-Hybrid Assays,Yeast Two-Hybrid Systems,n Hybrid System Techniques,n-Hybrid System Technique
D025941 Protein Interaction Mapping Methods for determining interaction between PROTEINS. Interaction Mapping, Protein,Interaction Mappings, Protein,Mapping, Protein Interaction,Mappings, Protein Interaction,Protein Interaction Mappings

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