用烟酰胺腺嘌呤二核苷酸（NAD）对白喉毒素片段A进行光亲和标记：第148位光产物的结构
Photoaffinity labeling of diphtheria toxin fragment A with NAD: structure of the photoproduct at position 148.
作者信息
Carroll S F, McCloskey J A, Crain P F, Oppenheimer N J, Marschner T M, Collier R J
出版信息
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7237-41. doi: 10.1073/pnas.82.21.7237.
Abstract
Irradiation of mixtures of diphtheria toxin fragment A and [carbonyl-14C]NAD with UV light (253.7 nm) is known to induce efficient transfer of the radiolabel to position 148, corresponding to glutamic acid in the unmodified protein. Here we report the structure of the photoproduct at position 148, as determined by chemical and photochemical methods, fast-atom-bombardment mass spectrometry, and nuclear magnetic resonance. The photoproduct [an alpha-amino-gamma-(6-nicotin-amidyl)butyric acid residue] contains the entire nicotinamide moiety of NAD linked via its number 6 carbon to the decarboxylated gamma-methylene carbon of Glu-148. No portion of the ADP-ribosyl group of NAD is present. These findings are consistent with the idea that Glu-148 lies at or near the catalytic center of diphtheria toxin.
摘要
已知用紫外线（253.7纳米）照射白喉毒素片段A与[羰基-¹⁴C]NAD的混合物，可使放射性标记有效地转移至148位，该位置对应于未修饰蛋白质中的谷氨酸。在此，我们报告通过化学和光化学方法、快原子轰击质谱法以及核磁共振确定的148位光产物的结构。该光产物[一个α-氨基-γ-（6-烟酰胺基）丁酸残基]包含NAD的整个烟酰胺部分，通过其6号碳与Glu-148脱羧的γ-亚甲基碳相连。NAD的ADP-核糖基团无任何部分存在。这些发现与Glu-148位于白喉毒素催化中心或其附近的观点一致。
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