通过皮秒紫外激光照射在重组核组蛋白、细胞核和全细胞中进行蛋白质-DNA交联。

Angelov D, Dimitrov S I, Russanova V R, Keskinova E, Pashev I G

Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia.

Nucleic Acids Res. 1988 May 25;16(10):4525-38. doi: 10.1093/nar/16.10.4525.

A picosecond UV laser was used to cross-link proteins to DNA in nuclei, whole cells and reconstituted nucleohistone. Irradiation of the nucleohistone resulted in crosslinking 15-20% of bound histones to DNA in a very short time (one or several picosecond pulses), the efficiency of crosslinking to single stranded DNA being higher than to double stranded DNA. All histones as well as high mobility group 1 proteins were identified in the covalently linked protein-DNA complexes upon irradiation of isolated nuclei and whole cells. A method is suggested for isolation of crosslinked material from cells and nuclei in amounts sufficient for further analysis. Experiments with reconstituted nucleohistones showed that upon irradiation at a constant dose the efficiency of crosslinking depended on the intensity of the light, thus suggesting a two-quantum process is involved in the reaction.

使用皮秒紫外激光将细胞核、全细胞和重组核组蛋白中的蛋白质与DNA交联。对核组蛋白进行照射会在极短时间内（一个或几个皮秒脉冲）使15%-20%的结合组蛋白与DNA交联，与单链DNA的交联效率高于双链DNA。在对分离的细胞核和全细胞进行照射后，共价连接的蛋白质-DNA复合物中鉴定出了所有组蛋白以及高迁移率族1蛋白。提出了一种从细胞和细胞核中分离交联物质的方法，其数量足以进行进一步分析。对重组核组蛋白的实验表明，在恒定剂量照射下，交联效率取决于光的强度，因此表明该反应涉及双量子过程。