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• W2019491472 abstract "The hydrogen exchange kinetics of the S-peptide in ribonuclease S can be measured by first tritiating the S-peptide in the absence of S-protein and then allowing it to recombine rapidly with S-protein. Afterwards the exchange reactions of this specific segment of ribonuclease S can be studied. The exchange kinetics of bound S-peptide are complex, indicating that different protons exchange at markedly different rates. The terminal exchange reaction, involving at least five highly protected protons, has been studied as a function of pH. At low concentrations of ribonuclease S the exchange kinetics become concentration-dependent, owing to the dissociation of the S-peptide. Although the fraction of free S-peptide is always very small, its rate of exchange is several orders of magnitude faster than that of bound S-peptide, and the concentration dependence of the exchange kinetics is readily measurable. It provides a highly sensitive method for determining small dissociation constants (KD). Values of KD ranging from 10−6m at pH 2.7, 0 °C, to 2 × 10−10m at pH 7.0, 0 °C, are reported here. Our value for KD at pH 7.0, 0 °C, confirms the data and extrapolation to 0 °C of Hearn et al. (1971). At high concentrations of ribonuclease S the terminal exchange reaction is independent of concentration. It probably results from a local unfolding reaction of the bound S-peptide. Above pH 4 the strong pH dependence of KD closely resembles that of the apparent equilibrium constant for this local unfolding reaction. The latter may be one step in the dissociation process and we present such a model for ribonuclease S dissociation. Measurement of concentration-dependent exchange kinetics should provide a useful method of determining small dissociation constants in other systems: for example, in studies of protein-nucleic acid interactions." @default.
• W2019491472 created "2016-06-24" @default.
• W2019491472 creator A5032058758 @default.
• W2019491472 creator A5067353361 @default.
• W2019491472 date "1976-08-01" @default.
• W2019491472 modified "2023-10-16" @default.
• W2019491472 title "Concentration-dependent hydrogen exchange kinetics of 3H-labeled S-peptide in ribonuclease S" @default.
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• W2019491472 doi "https://doi.org/10.1016/0022-2836(76)90101-7" @default.
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