Pautova L V, Laktionov P P, Rykova E Iu, Iakubov L A, Vlasov V V
Mol Biol (Mosk). 1994 Sep-Oct;28(5):1106-12.
Interaction of IgG molecules with oligonucleotides using reactive derivatives of p(T)16 bearing a 4-[(N-2-chloroethyl-N-methyl)amino]benzylamine residue at the 5'-terminal phosphate was investigated. The modified immunoglobulins were degraded with pepsin into Fab and Fc fragments and by incomplete CNBr hydrolysis into smaller peptides. It became obvious from the analysis of the peptides obtained that the oligonucleotides contacted with immunoglobulins at the antigen-binding Fab fragment of the molecule. The site of interaction is localized in the light-chain N-terminal fragment 178 amino acids long. These facts are in accordance with our previous data about the ability of a specific antigen to prevent the oligonucleotide-antibody interaction. On the contrary, an oligonucleotide covalently linked with immunoglobulin could not prevent the antigen-antibody reaction. Monoclonal Fab fragments modified with the alkylating p(T)16 derivative were found to interact with the specific antigen (human myoglobin) during affinity chromatography. This interaction deserves further investigation because of its importance in studying the fate of oligonucleotides in vivo.
研究了使用在5'-末端磷酸带有4-[(N-2-氯乙基-N-甲基)氨基]苄胺残基的p(T)16反应性衍生物，使IgG分子与寡核苷酸相互作用。修饰后的免疫球蛋白用胃蛋白酶降解为Fab和Fc片段，并用不完全的溴化氰水解为较小的肽段。从所得肽段的分析中可以明显看出，寡核苷酸在分子的抗原结合Fab片段处与免疫球蛋白接触。相互作用位点位于长度为178个氨基酸的轻链N末端片段中。这些事实与我们之前关于特定抗原阻止寡核苷酸-抗体相互作用能力的数据一致。相反，与免疫球蛋白共价连接的寡核苷酸不能阻止抗原-抗体反应。发现在亲和色谱过程中，用烷基化p(T)16衍生物修饰的单克隆Fab片段与特异性抗原（人肌红蛋白）相互作用。由于其在研究寡核苷酸在体内命运方面的重要性，这种相互作用值得进一步研究。
