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Suppression of problematic compound oligomerization by cosolubilization of nondetergent sulfobetaines.

Mizukoshi Y, Takeuchi K, Arutaki M, Takizawa T, Hanzawa H, Takahashi H, Shimada I - ChemMedChem (2015)

Bottom Line: In this study, solution NMR spectroscopy was used as a suitable technique to detect compound oligomers in equilibrium, and it was demonstrated that cosolubilization of nondetergent sulfobetaines (NDSBs) can largely suppress compound oligomerization and aggregation by shifting the equilibrium toward the monomers.The rotational correlation time of the aggregation-prone compound SKF86002 (1 mM) was substantially reduced from 0.31 to 0.23 ns by cosolubilization of 100 mM NDSB195.NDSB cosolubilization allowed us to perform successful structural and biochemical experiments with substantially fewer artifacts, which represents a strategy to directly resolve the problematic oligomerization and aggregation of compounds.

View Article: PubMed Central - PubMed

Affiliation: Biomedicinal Information Research Center (BIRC) and Molecular Profiling Research Center (Molprof), National Institute of Advanced Industrial Science and Technology (AIST), 2-3-26 Aomi, Koto-ku, Tokyo 135-0064 (Japan); Japan Biological Informatics Consortium (JBIC), 2-3-26 Aomi, Koto-ku, Tokyo 135-0064 (Japan).

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NDSB cosolubilization relieves the artificial inhibition of enzymatic activity. The aggregation-prone compounds, I4PTH (0.2 mm; ▪), clotrimazole (0.4 mm; ▪), and benzyl benzoate (1.0 mm; ▪), were added to the reaction solutions of the chymotrypsin assays with the indicated concentrations of NDSB256. The bars indicate the amount of product in the presence of the aggregation-prone compounds relative to that in the absence of the aggregation-prone compounds.
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fig05: NDSB cosolubilization relieves the artificial inhibition of enzymatic activity. The aggregation-prone compounds, I4PTH (0.2 mm; ▪), clotrimazole (0.4 mm; ▪), and benzyl benzoate (1.0 mm; ▪), were added to the reaction solutions of the chymotrypsin assays with the indicated concentrations of NDSB256. The bars indicate the amount of product in the presence of the aggregation-prone compounds relative to that in the absence of the aggregation-prone compounds.

Mentions: We found that cosolubilization of the NDSBs also prevented aggregation of other well-known aggregation-prone compounds. For tetraiodophenolphthalein (I4PTH),[4] the absorbance at λ=316 nm, which is specific to I4PTH, was increased in the presence of NDSB256 (Figure S5, Supporting Information). In addition, promiscuous inhibition of chymotrypsin activity by the aggregation-prone compounds benzyl benzoate, clotrimazole, and I4PTH was largely relieved by NDSB256 cosolubilization (Figure 5).[2,3] We performed a T1s/T1ns experiment with 1.0 mm benzyl benzoate cosolubilized with 100 mm NDSB256. In the experiment, the T1s/T1ns value of benzyl benzoate was 1.02. This value indicates that benzyl benzoate is not fully monomerized with 100 mm NDSB, owing to its stronger tendency for oligomerization and aggregation; this is consistent with the fact that the activity of chymotrypsin in the enzymatic assay was not fully recovered at this NDSB concentration (Figure 5). Notably, the T1s/T1ns value for benzyl benzoate was 1.00 with NDSB195 cosolubilization. The consistent T1s/T1ns values indicate that the T1s/T1ns experiment is not perturbed by the strong aromatic resonances originating from NDSB256, except for the case in which there is signal degeneracy.


Suppression of problematic compound oligomerization by cosolubilization of nondetergent sulfobetaines.

Mizukoshi Y, Takeuchi K, Arutaki M, Takizawa T, Hanzawa H, Takahashi H, Shimada I - ChemMedChem (2015)

NDSB cosolubilization relieves the artificial inhibition of enzymatic activity. The aggregation-prone compounds, I4PTH (0.2 mm; ▪), clotrimazole (0.4 mm; ▪), and benzyl benzoate (1.0 mm; ▪), were added to the reaction solutions of the chymotrypsin assays with the indicated concentrations of NDSB256. The bars indicate the amount of product in the presence of the aggregation-prone compounds relative to that in the absence of the aggregation-prone compounds.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4471626&req=5

fig05: NDSB cosolubilization relieves the artificial inhibition of enzymatic activity. The aggregation-prone compounds, I4PTH (0.2 mm; ▪), clotrimazole (0.4 mm; ▪), and benzyl benzoate (1.0 mm; ▪), were added to the reaction solutions of the chymotrypsin assays with the indicated concentrations of NDSB256. The bars indicate the amount of product in the presence of the aggregation-prone compounds relative to that in the absence of the aggregation-prone compounds.
Mentions: We found that cosolubilization of the NDSBs also prevented aggregation of other well-known aggregation-prone compounds. For tetraiodophenolphthalein (I4PTH),[4] the absorbance at λ=316 nm, which is specific to I4PTH, was increased in the presence of NDSB256 (Figure S5, Supporting Information). In addition, promiscuous inhibition of chymotrypsin activity by the aggregation-prone compounds benzyl benzoate, clotrimazole, and I4PTH was largely relieved by NDSB256 cosolubilization (Figure 5).[2,3] We performed a T1s/T1ns experiment with 1.0 mm benzyl benzoate cosolubilized with 100 mm NDSB256. In the experiment, the T1s/T1ns value of benzyl benzoate was 1.02. This value indicates that benzyl benzoate is not fully monomerized with 100 mm NDSB, owing to its stronger tendency for oligomerization and aggregation; this is consistent with the fact that the activity of chymotrypsin in the enzymatic assay was not fully recovered at this NDSB concentration (Figure 5). Notably, the T1s/T1ns value for benzyl benzoate was 1.00 with NDSB195 cosolubilization. The consistent T1s/T1ns values indicate that the T1s/T1ns experiment is not perturbed by the strong aromatic resonances originating from NDSB256, except for the case in which there is signal degeneracy.

Bottom Line: In this study, solution NMR spectroscopy was used as a suitable technique to detect compound oligomers in equilibrium, and it was demonstrated that cosolubilization of nondetergent sulfobetaines (NDSBs) can largely suppress compound oligomerization and aggregation by shifting the equilibrium toward the monomers.The rotational correlation time of the aggregation-prone compound SKF86002 (1 mM) was substantially reduced from 0.31 to 0.23 ns by cosolubilization of 100 mM NDSB195.NDSB cosolubilization allowed us to perform successful structural and biochemical experiments with substantially fewer artifacts, which represents a strategy to directly resolve the problematic oligomerization and aggregation of compounds.

View Article: PubMed Central - PubMed

Affiliation: Biomedicinal Information Research Center (BIRC) and Molecular Profiling Research Center (Molprof), National Institute of Advanced Industrial Science and Technology (AIST), 2-3-26 Aomi, Koto-ku, Tokyo 135-0064 (Japan); Japan Biological Informatics Consortium (JBIC), 2-3-26 Aomi, Koto-ku, Tokyo 135-0064 (Japan).

Show MeSH
Related in: MedlinePlus