Ligand and protein fishing with heat shock protein 90 coated magnetic beads.
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.
Marszałł
M P
MP
Gerontology Research Center, National Institute on Aging/NIH, Baltimore, Maryland 21224, USA.
Moaddel
R
R
Kole
S
S
Gandhari
M
M
Bernier
M
M
Wainer
I W
IW
D000255:Adenosine Triphosphate / Q000737:chemistry; D016227:Benzoquinones / Q000737:chemistry; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry* / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D047029:Lactams, Macrocyclic / Q000737:chemistry; D008024:Ligands; D008280:Magnetics / Q000379:methods*; D046912:Multiprotein Complexes / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D009675:Novobiocin / Q000737:chemistry; D011485:Protein Binding; D040901:Proteomics / Q000379:methods; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism
D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D055767:Immobilized Proteins; D047029:Lactams, Macrocyclic; D008024:Ligands; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; D009675:Novobiocin; D000255:Adenosine Triphosphate; C001277:geldanamycin