Whitlock J P, Stein A
J Biol Chem. 1978 Jun 10;253(11):3857-61.
HeLa chromatin core particles were digested with trypsin to excise the NH2-terminal histone regions. The resulting nucleoprotein complexes were dissociated in 2.5 M NaCl; the DNA and polypeptides were then allowed to reassemble by lowering the NaCl concentration. Eighty per cent of the DNA reassociated with the polypeptides. The reassembled nucleoprotein complexes sediment at 9.7 S, have a molecular elipticity at 280 nm of 3000 degrees cm2/dmol of PO4, and contain DNase I-susceptible sites at 10 nucleotide intervals. The pattern of products generated by cross-linking the polypeptides with dimethylsuberimidate is very similar to the pattern generated by cross-linking native core particles. The results indicate that histones which lack their HN2-terminal regions retain both the features necessary for correct protein-protein interactions and the ability to fold DNA into a nucleoprotein complex resembling the chromatin core particle.
将HeLa染色质核心颗粒用胰蛋白酶消化,以切除组蛋白的NH2末端区域。将所得的核蛋白复合物在2.5M NaCl中解离;然后通过降低NaCl浓度使DNA和多肽重新组装。80%的DNA与多肽重新结合。重新组装的核蛋白复合物在9.7S处沉降,在280nm处的分子椭圆率为3000度cm2/dmol的PO4,并且含有以10个核苷酸间隔的DNase I敏感位点。用亚胺基二甲酯使多肽交联产生的产物模式与天然核心颗粒交联产生的模式非常相似。结果表明,缺乏HN2末端区域的组蛋白既保留了正确的蛋白质-蛋白质相互作用所需的特征,又保留了将DNA折叠成类似于染色质核心颗粒的核蛋白复合物的能力。