模型系统和染色质中蛋白质与DNA的解离
Protein dissociation from DNA in model systems and chromatin.
作者信息
Shiffman M L, Maciewicz R A, Hu A W, Howard J C, Li H J
出版信息
Nucleic Acids Res. 1978 Sep;5(9):3409-26. doi: 10.1093/nar/5.9.3409.
Salt induced dissociation of protamine, poly(L-lysine) and poly(L-arginine) from DNA was measured by relative light scattering at theta = 90 degrees and/or centrifugation. Dissociation of histones from DNA was studied using relative light scattering and intrinsic tyrosine fluorescence. Protamine was dissociated from DNA at 0.15 M MgCl2 (ionic strength mu = 0.45) or 0.53 M NaCl (mu = 0.53) based on light scattering data and at approximately 0.2 M MgCl2 (mu = 0.6) or 0.6 M NaCl based on centrifugation data. NaCl induced dissociation of poly(Lys) or poly(Arg) from natural DNAs measured by light scattering did not depend on the guanine plus cytosine content. To dissociate poly(Arg) from DNA higher ionic strength using NaCl, MgCl2, or CaCl2, similar ionic strength using NaClo4, and lower ionic strength using Na2SO4 was needed then to dissociated poly(Lys). Both the decrease in light scattering and the enhancement of tyrosine fluorescence of chromatin occurred between 0.5 and 1.5 M NaCl when histones were dissociated.
通过在θ = 90°处的相对光散射和/或离心法测量盐诱导的鱼精蛋白、聚（L - 赖氨酸）和聚（L - 精氨酸）与DNA的解离。使用相对光散射和酪氨酸固有荧光研究组蛋白与DNA的解离。根据光散射数据，在0.15 M MgCl₂（离子强度μ = 0.45）或0.53 M NaCl（μ = 0.53）条件下，鱼精蛋白从DNA上解离；根据离心数据，在约0.2 M MgCl₂（μ = 0.6）或0.6 M NaCl条件下解离。通过光散射测量，NaCl诱导的聚（赖氨酸）或聚（精氨酸）从天然DNA上的解离不依赖于鸟嘌呤加胞嘧啶的含量。要使用NaCl、MgCl₂或CaCl₂以更高的离子强度使聚（精氨酸）从DNA上解离，使用高氯酸钠时需要相似的离子强度，而使用硫酸钠时需要更低的离子强度才能使聚（赖氨酸）解离。当组蛋白解离时，在0.5至1.5 M NaCl之间，染色质的光散射降低且酪氨酸荧光增强。
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