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Based on a model of smaller spheres packing on the surface of a larger sphere, the maximum binding stoichiometries of the nanoparticles (r ∼ 5 nm) with α-chymotrypsin (r ∼ 2.5 nm), cytochrome c (r ∼ 1.8 nm), and histone (r ∼ 2.1 nm) are evaluated as 28, 48, and 37, respectively. For the guideline regarding the calculation, see: Qi, K.; Ma, Q.-G.; Remsen, E. E.; Clark, C. G.; Wooley, K. L. J. Am. Chem. Soc. 2004, 126, 6599-6607.
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Based on a model of smaller spheres packing on the surface of a larger sphere, the maximum binding stoichiometries of the nanoparticles (r ∼ 5 nm) with α-chymotrypsin (r ∼ 2.5 nm), cytochrome c (r ∼ 1.8 nm), and histone (r ∼ 2.1 nm) are evaluated as 28, 48, and 37, respectively. For the guideline regarding the calculation, see: Qi, K.; Ma, Q.-G.; Remsen, E. E.; Clark, C. G.; Wooley, K. L. J. Am. Chem. Soc. 2004, 126, 6599-6607.
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The binding constants for NP-ChT interactions are around ∼ 10-fold lower than those obtained from enzyme activity assays (ref 17a), presumably because of the fact that in the latter case (i) the final solution contained 8% (v/v) of ethanol-DMSO (90:10) in the 5 mM phosphate buffer, (ii) 2 mM of N-succinyl-L-phenylalanine p-nitroanilide (SPNA) was presented as an enzyme substrate would be expected to interfere in the protein - NP interactions, and (iii) the protein concentrations for the ITC are ∼ 10-fold higher than those used in the activity assay, which would be expected to raise the ionic strength of the solution because of the polyelectrolyte nature of the protein.
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The binding constants for NP-ChT interactions are around ∼ 10-fold lower than those obtained from enzyme activity assays (ref 17a), presumably because of the fact that in the latter case (i) the final solution contained 8% (v/v) of ethanol-DMSO (90:10) in the 5 mM phosphate buffer, (ii) 2 mM of N-succinyl-L-phenylalanine p-nitroanilide (SPNA) was presented as an enzyme substrate would be expected to interfere in the protein - NP interactions, and (iii) the protein concentrations for the ITC are ∼ 10-fold higher than those used in the activity assay, which would be expected to raise the ionic strength of the solution because of the polyelectrolyte nature of the protein.
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NP concentrations were calculated on the basis of their average molecular weights (see Experimental Section and Supporting Information, Thus, the binding stoichiometrics between NPs and ChT differ slightly from previously reported stoichiometrics (ref 17a, where the NP concentrations were calibrated according to the UV absorbance of the gold core see: Link, S, Wang, Z.-L, El-Sayed, M. A. J. Phys. Chem. B 1999, 103, 3529-3533
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NP concentrations were calculated on the basis of their average molecular weights (see Experimental Section and Supporting Information). Thus, the binding stoichiometrics between NPs and ChT differ slightly from previously reported stoichiometrics (ref 17a), where the NP concentrations were calibrated according to the UV absorbance of the gold core (see: Link, S.; Wang, Z.-L.; El-Sayed, M. A. J. Phys. Chem. B 1999, 103, 3529-3533).
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TEM measurements on NP_Ala revealed an average particle size of 2.1 ± 0.4 nm (Figure S1). TGA revealed that the weight percentage of organic ligands in the NPs is 36% (Figure S2). Accordingly, the average molecular weight of the NP is estimated as 100 kDa, which was used for the preparation of NP solutions.
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TEM measurements on NP_Ala revealed an average particle size of 2.1 ± 0.4 nm (Figure S1). TGA revealed that the weight percentage of organic ligands in the NPs is 36% (Figure S2). Accordingly, the average molecular weight of the NP is estimated as 100 kDa, which was used for the preparation of NP solutions.
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