RESUMO
Heat shock protein 90alpha (Hsp90alpha) is a molecular chaperone that has been targeted for the development of new anticancer therapies. To date, co-immunoprecipitation (IP) has been primarily used to identify novel client proteins. We now report an alternative approach in which Hsp90alpha has been immobilized onto the surface of silica-based magnetic beads. The beads were used to isolate known Hsp90alpha ligands from a mixture containing ligands and nonligands. In addition, they were also used to isolated proteins from a mixture of proteins, as well as a cellular extract. The results indicate that the Hsp90alpha coated magnetic beads can be used to "fish" from complex chemical and biological mixtures for new lead drug candidates and client proteins.
Assuntos
Proteínas de Choque Térmico HSP90/química , Magnetismo/métodos , Complexos Multiproteicos/análise , Trifosfato de Adenosina/química , Benzoquinonas/química , Proteínas de Choque Térmico HSP90/metabolismo , Proteínas Imobilizadas/química , Proteínas Imobilizadas/metabolismo , Lactamas Macrocíclicas/química , Ligantes , Complexos Multiproteicos/isolamento & purificação , Complexos Multiproteicos/metabolismo , Novobiocina/química , Ligação Proteica , Proteômica/métodos , Proteínas Recombinantes/análise , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismoRESUMO
Human serum albumin, HSA, was immobilized onto the surface of silica-based magnetic beads. The beads were used to isolate known HSA ligands from a mixture containing ligands and nonligands. The separation was accomplished manually and was also automated. The results indicate that an automated "ligand-fishing" technique can be developed using magnetic beads containing an immobilized protein.
