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Figure 1.

Structure of compounds 1–7.

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Figure 2.

SPR analysis results.

Sensorgrams obtained by injecting different concentrations (from 0.020 to 1 µM) of 1 (A), 2 (B), 3 (C), 4 (D), 5 (E), 6 (F) 7 (G) and radiciol (H) on immobilized Hsp90.

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Table 1.

Thermodynamic and kinetic constants measured by SPR for the interaction between tested compounds and immobilized HSP90.

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Figure 3.

Inhibition of the ATPase activity of Hsp90 by different concentration of compounds 1–4.

Radicicol and 17-AAG were used as positive controls. Data are the mean of two independent experiments performed in triplicate and were analyzed by t test (Hsp90 vs Hsp90+ testing compound): *P<0.05, **P<0.005.

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Figure 4.

Aggregation kinetics of CS at 43°C determined by light scattering.

The spontaneous aggregation of CS at 43°C (♦) and the aggregation of CS at 43°C in the presence of 0.075 µM Hsp90 (Δ) or of 0.075 µM Hsp90 and 0.3 µM compound 1 (▪), 0.3 µM compound 2 (•), 0.3 compound 3 (□), or 0.3 µM compound 4 (○) are shown.

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Figure 5.

Schematic representation of the results obtained from limited proteolysis experiments.

The preferential cleavage sites detected on recombinant Hsp90, and on the Hsp90/1 complex are in black. The Hsp90 N-terminal domain is highlighted in light grey, the middle domain is boxed and the C-terminal domain is highlighted in grey.

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Figure 6.

Docking calculation results.

Three dimensional models (A and B) of (+)-lentiginosine (1, yellow) and (−)-lentiginosine (2, green) with HSP90. The target molecule is depicted by sky blue ribbon and the crucial amino acids by cpk (by atom type: C, purple; O, red; N, dark blue, H, white).

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