TY - JOUR
T1 - Quantitative analysis of Hsp90-client interactions reveals principles of substrate recognition
AU - Taipale, Mikko
AU - Krykbaeva, Irina
AU - Koeva, Martina
AU - Kayatekin, Can
AU - Westover, Kenneth D.
AU - Karras, Georgios I.
AU - Lindquist, Susan
N1 - Funding Information:
We would like to thank D.F. Jarosz, L. Whitesell, and M. Mendillo for valuable comments, K. Salehi-Ashtiani and M. Vidal (Dana-Farber Cancer Institute) for the human ORFeome collection, D. Proia (Synta Pharmaceuticals) for ganetespib, and N.S. Gray (Dana-Farber Cancer Institute) for DPH. M.T. was supported by the Academy of Finland, Sigrid Juselius Foundation and the Human Frontier Science Program. G.I.K. was supported by an EMBO Long-Term Fellowship and the Human Frontier Science Program. S.L. is an investigator of the Howard Hughes Medical Institute. Support for this study was also provided by the NIH Genomics Based Drug Discovery-Driving Medical Projects grant UL1-DE019585, administratively linked to NIH grants RL1-GM084437, RL1-CA133834, and RL1-HG004671. M.T. and S.L. planned the project and designed the experiments. M.T. developed and validated LUMIER with BACON assay and performed all the experiments with the help of I.K. G.I.K. assayed HSP90 interactions against E3 ligases. K.D.W. and M.T. expressed and purified recombinant ABL, and C.K. and M.T. analyzed ABL with circular dichroism. M.T. and M.K. performed computational analyses of HSP90 interactions. M.T. and S.L. wrote the paper, with comments from all the coauthors.
PY - 2012/8/31
Y1 - 2012/8/31
N2 - HSP90 is a molecular chaperone that associates with numerous substrate proteins called clients. It plays many important roles in human biology and medicine, but determinants of client recognition by HSP90 have remained frustratingly elusive. We systematically and quantitatively surveyed most human kinases, transcription factors, and E3 ligases for interaction with HSP90 and its cochaperone CDC37. Unexpectedly, many more kinases than transcription factors bound HSP90. CDC37 interacted with kinases, but not with transcription factors or E3 ligases. HSP90::kinase interactions varied continuously over a 100-fold range and provided a platform to study client protein recognition. In wild-type clients, HSP90 did not bind particular sequence motifs, but rather associated with intrinsically unstable kinases. Stabilization of the kinase in either its active or inactive conformation with diverse small molecules decreased HSP90 association. Our results establish HSP90 client recognition as a combinatorial process: CDC37 provides recognition of the kinase family, whereas thermodynamic parameters determine client binding within the family.
AB - HSP90 is a molecular chaperone that associates with numerous substrate proteins called clients. It plays many important roles in human biology and medicine, but determinants of client recognition by HSP90 have remained frustratingly elusive. We systematically and quantitatively surveyed most human kinases, transcription factors, and E3 ligases for interaction with HSP90 and its cochaperone CDC37. Unexpectedly, many more kinases than transcription factors bound HSP90. CDC37 interacted with kinases, but not with transcription factors or E3 ligases. HSP90::kinase interactions varied continuously over a 100-fold range and provided a platform to study client protein recognition. In wild-type clients, HSP90 did not bind particular sequence motifs, but rather associated with intrinsically unstable kinases. Stabilization of the kinase in either its active or inactive conformation with diverse small molecules decreased HSP90 association. Our results establish HSP90 client recognition as a combinatorial process: CDC37 provides recognition of the kinase family, whereas thermodynamic parameters determine client binding within the family.
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U2 - 10.1016/j.cell.2012.06.047
DO - 10.1016/j.cell.2012.06.047
M3 - Article
C2 - 22939624
AN - SCOPUS:84865695733
SN - 0092-8674
VL - 150
SP - 987
EP - 1001
JO - Cell
JF - Cell
IS - 5
ER -