A New Functional Model for Prediction of Chaperone Activity of the Recombinant M. tb Acr (α-Crystallin) Using Insulin as Substrate

Figure 5

Molecular interaction of (G) and (H) samples of Acr without pre-heat treatments. (a) Plot of molecules of β-sheets versus % inhibition for (G) samples without pre-heat treatment: the % of β-sheets (45.5%) obtained from the CD analysis (Table 4) was multiplied by the total number of Acr molecules at 4 different concentrations of Acr 6, 7, 9, and 12 μM to estimate the actual number of β-sheets interacting with insulin. The total amount of Acr in the reaction was recalculated based on the Native-PAGE data and the ratio of oligomers (40% of 24 mers and 15% of 9, 10, 11, and 12 mers) (Table 1). This calculation was repeated for all (G) samples. (b) Plot of α-helices versus % inhibition for (G) samples without pre-heat treatment: the % of α-helices for (G) samples was 9.6% (Table 4), and this was multiplied with the 4 different concentrations of Acr 6, 7, 9, and 12 μM to estimate the actual number of α-helices interacting with insulin. (c) Plot of random coils versus % inhibition for (G) samples without pre-heat treatment: the % of random coils of (G) samples (34.4%) (Table 4) was multiplied with 4 different concentrations of Acr 6, 7, 9, and 12 μM to estimate the actual number of random coils interacting with insulin. The total amount of Acr in the reaction was recalculated based on the Native-PAGE data and the ratio of oligomers (40% of 24 mers and 15% of 9, 10, 11, and 12 mers) (Table 1). This calculation was repeated for all (G) samples. (d) Plot of molecules of β-sheets versus % inhibition for (H) samples: the % of β-sheets (2.5%) (Table 3) was multiplied by the total number of Acr molecules at 4 different concentrations of Acr 5.5, 11, 18, and 30 μM to estimate the actual number of β-sheets interacting with insulin. The total amount of Acr in the reaction was recalculated based on Native-PAGE data and the ratio of oligomers (70% of 9 mers and 10% of 10, 11, and 12 mers) (Table 1). This process was repeated for all (H) samples. (e) Plot of α-helices versus % inhibition for (H) samples without pre-heat treatment: the % of α-helices for the (H) samples was 26% (Table 3), and this was multiplied with the 4 different concentrations of Acr 5.5, 11, 18, and 30 μM to estimate the actual number of α-helices interacting with insulin. The total amount of Acr in the reaction was recalculated based on Native-PAGE data and the ratio of oligomers (70% of 9 mers and 10% of 10, 11, and 12 mers) (Table 1). This process was repeated for all (H) samples. (f) Plot of random coils versus % inhibition for (H) samples without pre-heat treatment: the % of random coils for the (H) samples was 36.2% (Table 3), and this was multiplied with 4 different concentrations of Acr 5.5, 11, 18, and 30 μM to estimate the actual number of turns interacting with insulin. The total amount of Acr in the reaction was recalculated based on Native-PAGE data and the ratio of oligomers (70% of 9 mers and 10% of 10, 11, and 12 mers) (Table 1). This process was repeated for all (H) samples.

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