Rippin Thomas M, Freund Stefan M V, Veprintsev Dmitry B, Fersht Alan R
Cambridge University Chemical Laboratory and Cambridge Centre for Protein Engineering, MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
J Mol Biol. 2002 May 31;319(2):351-8. doi: 10.1016/S0022-2836(02)00326-1.
We present an analysis by NMR of a 58 kDa complex of the core domain of the tumour suppressor p53 with DNA that complements and extends the crystal structure analysis. Binding of specific DNA caused significant chemical shifts of residues on the DNA-binding interface that translated into the beta-sheet of the protein. Binding of non-specific DNA caused weak but qualitatively the same shifts, corresponding to weaker binding interactions. The observed chemical shift differences correlate with frequency of cancer-inducing mutations, suggesting that the affected residues contribute to the stability of p53 core domain-DNA complex. We also identified two affected regions on the surface of the protein: helix 1 (residues V173-C182) plus G244 and residues L114-T118, which may represent a dimerisation interface.
我们通过核磁共振（NMR）对肿瘤抑制蛋白p53核心结构域与DNA形成的58 kDa复合物进行了分析，该分析补充并扩展了晶体结构分析。特异性DNA的结合导致DNA结合界面上的残基发生显著化学位移，这些位移延伸至蛋白质的β折叠结构。非特异性DNA的结合导致微弱但性质相同的位移，这对应于较弱的结合相互作用。观察到的化学位移差异与致癌突变频率相关，表明受影响的残基有助于p53核心结构域 - DNA复合物的稳定性。我们还在蛋白质表面确定了两个受影响的区域：螺旋1（残基V173 - C182）加上G244以及残基L114 - T118，它们可能代表一个二聚化界面。
