关于使用紫外线研究蛋白质 - DNA 交联 - Suppr

Celis J E, Fink M, Kaltoft K

Nucleic Acids Res. 1976 Apr;3(4):1065-71. doi: 10.1093/nar/3.4.1065.

Irradiation of Ehrlich ascites chromatin with ultraviolet light (u.v.) leads to protein-DNA crosslinking as determined by CsCl isopycnic ultra-centrifugation and SDS-polyacrylamide gel electrophoresis. At the most 4.5% of the chromatin proteins labelled with (14C)-lysine and (14C)-arginine can be crosslinked to DNA at u.v. doses between 3.6 X 10(4) to 10.8 X 10(4) ergs/mm2. We find however that the crosslinking reaction is hindered by protein breakage induced by u.v. light of wave length of less than 2950 A. Our results point out that caution must be used in the interpretation of studies on protein-nucleic acid interactions using u.v. light.

用氯化铯等密度超速离心法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测定，用紫外线（uv）照射艾氏腹水染色质会导致蛋白质与DNA交联。在3.6×10⁴至10.8×10⁴尔格/平方毫米的紫外线剂量下，最多4.5%用（¹⁴C）-赖氨酸和（¹⁴C）-精氨酸标记的染色质蛋白可与DNA交联。然而，我们发现，波长小于2950埃的紫外线诱导的蛋白质断裂会阻碍交联反应。我们的结果指出，在解释使用紫外线进行的蛋白质-核酸相互作用研究时必须谨慎。