Johnsson Nils
Institut für Toxikologie und Genetik, Forschungszentrum Karlsruhe, Karlsruhe, Germany.
FEBS Lett. 2002 Nov 6;531(2):259-64. doi: 10.1016/s0014-5793(02)03533-0.
Many mutations in the human tumor suppressor p53 affect the function of the protein by destabilizing the structure of its DNA binding domain. To monitor the effects of those mutations in vivo the stability of the DNA binding domain of p53 and some of its mutants was investigated with the split-ubiquitin (split-Ub) method. The split-Ub-derived in vivo data on the relative stability of the mutants roughly correlate with the quantitative data from in vitro denaturation experiments as reported in the literature. A variation of this assay allows visualizing the difference in stability between the wild-type p53 core and the mutant p53(V143A) by a simple growth assay.
人类肿瘤抑制蛋白p53中的许多突变会破坏其DNA结合结构域的结构，从而影响该蛋白的功能。为了在体内监测这些突变的影响，采用分裂泛素（split-Ub）方法研究了p53及其一些突变体的DNA结合结构域的稳定性。来自分裂泛素的关于突变体相对稳定性的体内数据与文献中报道的体外变性实验的定量数据大致相关。该检测方法的一种变体能够通过简单的生长检测直观呈现野生型p53核心与突变型p53（V143A）之间的稳定性差异。
