小牛胸腺组蛋白组分在水溶液中与8-苯胺基萘-1-磺酸的相互作用。

Interactions of calf-thymus histone fractions in aqueous solution with 8-anilinonaphthalene-1-sulphonic acid.

作者信息

Laurence D J

出版信息

Biochem J. 1966 May;99(2):419-26. doi: 10.1042/bj0990419.

DOI:10.1042/bj0990419

PMID:5944245

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1265011/

Abstract

The interactions of histone fractions with 8-anilinonaphthalene-1-sulphonic acid were investigated by fluorimetry and spectrofluorimetry and the results were interpreted with the aid of equilibrium-dialysis techniques. 2. Characteristic differences were found between the various histone fractions, and with fractions F3 and F2a the binding was found to be salt-dependent. 3. Evidence was obtained indicating a slow change of the physical state of fractions F3 and F2a in the presence of salt, and the binding by these two fractions in the presence of salt was greater by an order of magnitude than by fractions F1 and F2b. 4. Conditions favouring binding were also those favouring histone aggregation; SO(4) (2-) ions activated binding at a lower concentration than Cl(-) ions; urea, guanidinium ions and high concentrations of I(-) ions were inhibitory to binding. 5. After histones had been kept in the presence of salt for a long time the reversal of interaction on decreasing the salt concentration was incomplete. 6. The inhibition of binding by fraction F2a in the presence of urea or fraction F2b depended on the time sequence of addition of the reagents. 7. Artificial nucleoproteins made by precipitating DNA with the histone fractions in neutral 0.14m-sodium chloride showed the same order of interaction as was found for the fractions in solution. 8. Comparison of the binding by fraction F2a with that by bovine plasma albumin showed that in both cases there were a large number of weakly binding sites but that fraction F2a lacked the small number of strongly binding sites found in albumin. No slow change of binding in the presence of salt was found for albumin. 9. Binding by fraction F2b increased the affinity of the protein for further molecules of the adsorbate. 10. The results are discussed in relation to the close relationship between binding and aggregation and the possible role of non-polar interactions as determined by the balance between polar and non-polar amino acids in the histone fractions.

摘要

采用荧光分析法和荧光分光光度法研究了组蛋白组分与8-苯胺基萘-1-磺酸的相互作用，并借助平衡透析技术对结果进行了解释。2. 发现不同组蛋白组分之间存在特征差异，对于F3和F2a组分，发现其结合作用依赖于盐。3. 有证据表明，在盐存在的情况下，F3和F2a组分的物理状态会缓慢变化，并且这两个组分在盐存在时的结合作用比F1和F2b组分大一个数量级。4. 有利于结合的条件也是有利于组蛋白聚集的条件；SO(4) (2-)离子在比Cl(-)离子更低的浓度下就能激活结合；尿素、胍离子和高浓度的I(-)离子对结合有抑制作用。5. 组蛋白在盐存在的情况下长时间保存后，降低盐浓度时相互作用的逆转并不完全。6. 在尿素存在下F2a组分或F2b组分对结合的抑制作用取决于试剂添加的时间顺序。7. 通过在中性0.14m氯化钠中用组蛋白组分沉淀DNA制备的人工核蛋白，其相互作用顺序与溶液中组分的相互作用顺序相同。8. 比较F2a组分与牛血浆白蛋白的结合作用表明，在这两种情况下都有大量弱结合位点，但F2a组分缺乏白蛋白中发现的少量强结合位点。白蛋白在盐存在时未发现结合作用的缓慢变化。9. F2b组分的结合增加了蛋白质对吸附物进一步分子的亲和力。10. 结合结果与结合和聚集之间的密切关系以及组蛋白组分中极性和非极性氨基酸平衡所决定的非极性相互作用的可能作用进行了讨论。