Schnackerz Klaus D, Andi Babak, Cook Paul F
Physiologische Chemie I, Theodor-Boveri-Institut für Biowissenschaften, Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Biochim Biophys Acta. 2011 Nov;1814(11):1447-58. doi: 10.1016/j.bbapap.2011.02.001. Epub 2011 Feb 25.
In this review it is demonstrated that (31)P NMR spectroscopy can be used to elucidate information about the microenvironment around the phosphate group of enzyme-bound pyridoxal 5'-phosphate (PLP). The following information can be obtained for all PLP-dependent enzymes: 1) the protonation state of the 5'-phosphate and its exposure to solvent, and 2) tightness of binding of the 5'-phosphate. In addition, the 5-phosphate can report on the protonation state of the Schiff base lysine in some enzymes. Changes in the 5'-phosphate chemical shift can be used to determine changes in tightness of binding of the phosphate as the reaction pathway is traversed, providing information on the dynamics of the enzyme. (31)P NMR spectroscopy is thus an important probe of structure, dynamics and mechanism in native and site-directed mutations of PLP-dependent enzymes. Examples of all of the above are provided in this review. This article is part of a Special Issue entitled: Pyridoxal Phospate Enzymology.
本综述表明，磷-31核磁共振光谱可用于阐明有关与酶结合的磷酸吡哆醛-5'-磷酸（PLP）磷酸基团周围微环境的信息。对于所有依赖PLP的酶，可获得以下信息：1）5'-磷酸的质子化状态及其与溶剂的接触情况，以及2）5'-磷酸的结合紧密程度。此外，在某些酶中，5-磷酸可反映席夫碱赖氨酸的质子化状态。随着反应途径的进行，5'-磷酸化学位移的变化可用于确定磷酸结合紧密程度的变化，从而提供有关酶动力学的信息。因此，磷-31核磁共振光谱是研究PLP依赖酶的天然形式和定点突变的结构、动力学及作用机制的重要探针。本综述提供了上述所有方面的实例。本文是名为“磷酸吡哆醛酶学”的特刊的一部分。
