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. Author manuscript; available in PMC: 2021 Sep 23.
Published in final edited form as: Nat Struct Mol Biol. 2009 Oct 18;16(11):1141–1147. doi: 10.1038/nsmb.1682

Figure 6.

Figure 6

β-strand 8 mutation alters the conformational state of the charged linker. (a–c) COS7 cells were transfected with indicated Flag-HSP90 constructs. (a) Cells were lysed and proteins were immunoprecipitated by Flag antibody–conjugated agarose. Immunoprecipitates were digested with TCPK-treated trypsin and detected by immunoblotting with N-terminal antibody to HSP90. Molecular weight (kDa) is shown on the left. (b) Cells were lysed, and reactivity of natively folded HSP90 proteins against N terminus or charged-linker antibody was determined by ELISA. Error bars represent s.d. from the mean. (c) Cells were lysed, and reactivity of denatured HSP90 proteins against N-terminal or charged-linker antibody was determined by immunoblotting. CL, charged-linker antibody; N, N-terminus antibody.