Hydrogen-deuterium exchange rate between a peptide group and an aqueous solvent as determined by a stopped-flow ultraviolet spectrophotometry.

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概要

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• When an <SUP>1</SUP>H<SUB>2</SUB>O solution of <I>N</I>-methylacetamide was rapidly mixed with <SUP>2</SUP>H<SUB>2</SUB>O, a time-dependent decrease of the ultraviolet absorbance at 227 nm was observed. This was not the case with <I>N</I>,<I>N</I>-dimethylacetamide. Hence, the time-dependent decrease in absorbance was attributed to be caused by the deuteration (NH→ND) reaction of <I>N</I>-methylacetamide. This provides an appropriate method for determining deuteration rate of the peptide group in a polypeptide or in a protein. By this method the deuteration rate of poly-D,L-alanine has been examined in the pH=5—8 and temperature=10—40 °C range. In this range, the deuteration was found to proceed always as a single first-order reaction, and the rate constant <I>k</I><SUB>e</SUB> is expressed within an experimental error as <I>k</I><SUB>e</SUB>=6.4×10<SUP>(6+<I>pH</I>−3850⁄<I>T</I>)</SUP> This is to be used as a standard rate-constant value in estimating the attenuation factor γ<I><SUB>j</SUB></I>=<I>k<SUB>j</SUB></I>⁄<I>k</I><SUB>e</SUB> of a given peptide group (<I>j</I>) in a protein molecule whose rate constant value is <I>k<SUB>j</SUB></I>.

• 公益社団法人 日本化学会の論文

著者

• Takahashi Tetsuo

  Faculty Of Pharmaceutical Sciences Hokkaido University

• Nakanishi Mamoru

  Faculty Of Pharmaceutical Science University Of Tokyo

• Tsuboi Masamichi

  Faculty of Parmaceutical Sciences, The University of Tokyo

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