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Merck
  • Detection of ligand binding to proteins through observation of hydration water.

Detection of ligand binding to proteins through observation of hydration water.

The journal of physical chemistry. B (2012-11-16)
Saroj Kumar, Nadejda Eremina, Andreas Barth
摘要

Drug development is impeded by the need to design for each drug target a test that detects the binding of drug candidate molecules to the target protein. Therefore, a general method to detect ligand binding is highly desirable. Here, we present an observation toward developing such a method, which is based on monitoring a change in water absorption by infrared spectroscopy. Infrared spectroscopy has high sensitivity for water, and changes in its hydrogen bond pattern can be observed. We studied absorption changes of water upon the addition of phosphenolpyruvate or Mg(2+) to pyruvate kinase. In each case, there is a decrease in the absorption of water in the 3000-3100 cm(-1) region on the low wavenumber side of the OH stretching vibration when a ligand binds to the protein. Our results suggest that the weaker water absorption is due to the release of protein-bound water into bulk water during ligand binding. This observation has high potential for drug development as well as for basic research because it can lead to a general method for detecting molecular association events that (i) is label-free, (ii) works with both binding partners being in aqueous solution, and (iii) is based on a universal process that takes place in all binding events.

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Sigma-Aldrich
丙酮酸激酶 来源于兔肌肉, Type III, lyophilized powder, 350-600 units/mg protein
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丙酮酸激酶 来源于兔肌肉, Type II, ammonium sulfate suspension, 350-600 units/mg protein
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丙酮酸激酶 来源于兔肌肉, Type VII, buffered aqueous glycerol solution, 350-600 units/mg protein
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丙酮酸激酶 来源于嗜热脂肪芽胞杆菌, Type VIII, lyophilized powder, 100-300 units/mg protein