Leibniz Institute on Aging - Fritz Lipmann Institute, Beutenbergstr. 11, D-07745 Jena, Germany.
Institute for Organic Chemistry and Macromolecular Chemistry, Friedrich Schiller University, Humboldtstr. 10, D-07743 Jena, Germany.
J Magn Reson. 2019 Nov;308:106561. doi: 10.1016/j.jmr.2019.07.048. Epub 2019 Jul 16.
The N-terminal segment of human cystathionine-β-synthase (CBS(1-40)) constitutes an intrinsically disordered protein stretch that transiently interacts with heme. We illustrate that the HCBCACON experimental protocol provides an efficient alternative approach for probing transient interactions of intrinsically disordered proteins with heme in situations where the applicability of the conventional [H, N]-HSQC experiment may be limited. This experiment starting with the excitation of protein side chain protons delivers information about the proline residues and thereby makes it possible to use these residues in interaction mapping experiments. Employing this approach in conjunction with site-specific mutation we show that transient heme binding is mediated by the Cys-Pro motif of CBS(1-40).
人胱硫醚-β-合酶（CBS(1-40)）的 N 端片段构成一个内在无规的蛋白质延伸段，它与血红素发生瞬时相互作用。我们表明，HCBCACON 实验方案为探测内在无序蛋白质与血红素的瞬时相互作用提供了一种有效的替代方法，这种方法在常规 [H, N]-HSQC 实验的适用性可能受到限制的情况下尤其有用。这个实验从激发蛋白质侧链质子开始，提供关于脯氨酸残基的信息，从而使这些残基能够在相互作用映射实验中使用。我们采用这种方法结合定点突变，表明瞬时血红素结合是由 CBS(1-40)的 Cys-Pro 基序介导的。
