Garner M M, Rau D C
Laboratory of Theoretical and Physical Biology, NICHD, National Institutes of Health, Bethesda, MD 20892, USA.
EMBO J. 1995 Mar 15;14(6):1257-63. doi: 10.1002/j.1460-2075.1995.tb07109.x.
Water release coupled to the association of gal repressor with DNA is measured from the sensitivity of the binding constant to the solution osmotic pressure, using neutral solutes that are typically excluded from polar protein and DNA surfaces. Differences in water release for binding of repressor to different sequences are linked with differences in specificity and binding energies. With sucrose, the specific binding of repressor to operator sequences is accompanied by the release of 130 water molecules. No water release is seen for the weak, non-specific binding of repressor to poly(dI-dC).(dI-dC). A difference in the release of six water molecules is seen even for the binding of gal repressor to two different operator sequences that differ in affinity by only a factor of two.
通过结合常数对溶液渗透压的敏感性来测量与半乳糖阻遏物和DNA结合相关的水释放,使用通常被极性蛋白质和DNA表面排斥的中性溶质。阻遏物与不同序列结合时水释放的差异与特异性和结合能的差异相关。对于蔗糖,阻遏物与操纵序列的特异性结合伴随着130个水分子的释放。阻遏物与聚(dI-dC)·(dI-dC)的弱非特异性结合未观察到水释放。即使半乳糖阻遏物与亲和力仅相差两倍的两个不同操纵序列结合,也观察到了六个水分子释放的差异。