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Ultrasonic absorption studies of protein-buffer interactions

Determination of equilibrium parameters of titratable groups

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Abstract

The acoustic absorption of protein solutions in the presence of phosphate and other buffering ions has been studied in the physiological pH range. Buffers containing hydroxyl residues as titratable groups cause a pronounced increase of protein sound absorption, which is attributed to relaxation processes of proton transfer reactions between buffer ions and accessible imidazole and α-amino groups of the protein surface. Amino group based buffers like Good's buffers do not induce additional sound absorption. Measurement of the ultrasonic absorption as a function of pH and of buffer concentration, and corresponding parameter fitting of the equation describing proton transfer relaxation processes has been used to evaluate equilibrium parameters. For the imidazole group of the amino acid histidine a pK value of 6.22 and for the imidazole group of the protein lysozyme a pK value of 5.71 have been determined. In hemoglobin the ligand-linked pK changes have been monitored by recording ultrasonic titration curves.

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Correspondence to K. D. Jürgens.

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Jürgens, K.D., Baumann, R. Ultrasonic absorption studies of protein-buffer interactions. Eur Biophys J 12, 217–222 (1985). https://doi.org/10.1007/BF00253848

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Key words

  • Ultrasonic absorption
  • equilibrium constants
  • histidine
  • lysozyme
  • hemoglobin