Knorre D G, Bichenkova E V, Koval' V V, Alekseev P V, Knorre V D, Nordhoff E, Godovikova T S
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division, Russian Academy of Sciences, Novosibirsk, Russia.
Bioorg Khim. 1998 Sep;24(9):663-9.
A new approach to the study of the interaction of amino acid side chains with photoreactive aryl azides was proposed. This approach was based on the drawing together of the reacting groups by the attachment of the reacting compounds to complementary oligonucleotides. Cystamine, histamine, and 1,6-hexamethylenediamine mimicking the cystine, histidine, and lysine residues, respectively, were attached to the 3'-terminal phosphate of the oligonucleotide GGTATCp through a phosphamide bond and used as the targets for photomodification. Derivatives of the oligonucleotide pGATACCAA with the fragment N3C6H4NH- attached directly to its 5'-end by a phosphamide bond or through the spacer -(CH2)nNH- (where n is 2, 4, and 6) were used as photoreagents. Their derivatives containing the same spacer and the N3C6F4CO-NH(CH2)3NH- or 2-N3,5-NO2-C6H3CO-NH(CH2)3NH- residues were also used. The duplexes were photomodified by irradiation with 300-350 nm wavelength light. The maximal yields of the photo-cross-linking were from 22 to 68%. The reagents containing p-azidoaniline residue were found to be the most effective toward the targets. The maximum yields of the photomodification products modeling the side chains of cysteine and lysine were found to vary from 40 to 67% and to depend on the length and the structure of the spacers used. The duplex with the target bearing the imidazole residue (the histidine model) manifested a yield decreased to 25%. This fact was in a good agreement with the data of computer modeling that indicated an unfavorable mutual displacement of the imidazole residue and the photoreactive group.
提出了一种研究氨基酸侧链与光反应性芳基叠氮化物相互作用的新方法。该方法基于通过将反应化合物连接到互补寡核苷酸上，使反应基团靠近。分别模拟胱氨酸、组氨酸和赖氨酸残基的胱胺、组胺和1,6 - 己二胺通过磷酰胺键连接到寡核苷酸GGTATCp的3'-末端磷酸上，并用作光修饰的靶标。寡核苷酸pGATACCAA的衍生物通过磷酰胺键或通过间隔基-(CH2)nNH-（其中n为2、4和6）直接连接到其5'-末端，用作光试剂。还使用了含有相同间隔基和N3C6F4CO-NH(CH2)3NH-或2-N3,5-NO2-C6H3CO-NH(CH2)3NH-残基的衍生物。通过用波长300 - 350nm的光照射对双链体进行光修饰。光交联的最大产率为22%至68%。发现含有对叠氮苯胺残基的试剂对靶标最有效。模拟半胱氨酸和赖氨酸侧链的光修饰产物的最大产率在40%至67%之间变化，并取决于所用间隔基的长度和结构。带有咪唑残基（组氨酸模型）的靶标的双链体产率降至25%。这一事实与计算机模拟数据非常吻合，该数据表明咪唑残基和光反应基团的相互位移不利。
