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

Table 5

Molecular interaction studies. Calculations of molecular level interaction of Acr (G), (H), and (G) with pre-heat treatment (concentrations of 12 μM, 30 μM, and 1 μM, respectively) are based on the following assumptions: reaction volume 0.25 ml, insulin molecular weight of B chain 3 kDa, and Acr molecular weight 18 kDa. The total amount of Acr in the reaction is recalculated based on the Native-PAGE data and the ratio of oligomers assuming an error estimation of ±10 % (Table 1).

	Mol. wt. (kDa)	Amount (µg)	Molecules of insulin B chain alone	% of insulin B chain covered
Insulin	6	174	8.73 × 10^15
Acr	18
(G) (12 µM)	18	54	1.8 × 10^15
40% of 24 mers	288	21.6	3.011 × 10^13 ± 0.301 × 10^13
15% of 9 mers	162	8.1	2.98 × 10^13 ± 0.298 × 10^13
15% of 10 mers	180	8.1	2.7 × 10^13 ± 0.27 × 10^13
15% of 11 mers	198	8.1	2.48 × 10^13 ± 0.248 × 10^13
15% of 12 mers	216	8.1	2.28 × 10^13 ± 0.228 × 10^13
Total Acr molecules			1.35 × 10^14	1.54
(H) (30 µM)		135	4.52 × 10^15 ± 0.452 × 10^15
70% of 9 mers	162	94.5	3.5 × 10^14 ± 0.35 × 10^14
10% of 10 mers	180	13.5	4.39 × 10^13 ± 0.439 × 10^13
10% of 11 mers	198	13.5	4.13 × 10^13 ± 0.413 × 10^13
10% of 12 mers	216	13.5	3.76 × 10^13 ± 0.376 × 10^13
Total Acr molecules			4.73 × 10^14 ± 0.473 × 10^14	6
(G) (1 µM) with pre-heat treatment	18	4.5	1.50 × 10^14
40% of 24 mers	288	1.8	2.49 × 10^12 ± 0.249 × 10^12
15% of 9 mers	162	0.68	2.53 × 10^12 ± 0.253 × 10^12
15% of 10 mers	180	0.68	2.28 × 10^12 ± 0.228 × 10^12
15% of 11 mers	198	0.68	2.07 × 10^12 ± 0.207 × 10^12
15% of 12 mers	216	0.68	1.88 × 10^12 ± 0.188 × 10^12
Total Acr molecules			1.13 × 10^13	0.13