Warner D R, Hoffman J L
University of Louisville, Department of Biochemistry, Kentucky 40292.
Anal Biochem. 1991 Jun;195(2):265-8. doi: 10.1016/0003-2697(91)90327-p.
A method is described for the synthesis and purification of the photoaffinity label Se-(8-azidoadenosyl)[75Se]selenomethionine. This photoaffinity label can be used to specifically and covalently label the S-adenosylmethionine binding site of proteins that use this cofactor, as exemplified by labeling of thioether methyltransferase. By utilizing the gamma-emitting isotope of selenium, Se-(8-azidoadenosyl)[75Se]selenomethionine eliminates the need for the impregnation of acrylamide gels with fluorographic enhancers and dilution of liquid samples into scintillation cocktails, as is required with the commonly used methyl-3H-labeled and 35S-labeled S-(8-azidoadenosyl)methionine.
描述了一种用于合成和纯化光亲和标记物硒-(8-叠氮腺苷)[75Se]硒代蛋氨酸的方法。这种光亲和标记物可用于特异性地和共价地标记使用该辅因子的蛋白质的S-腺苷甲硫氨酸结合位点,硫醚甲基转移酶的标记就是一个例子。通过利用硒的γ发射同位素,硒-(8-叠氮腺苷)[75Se]硒代蛋氨酸无需像常用的甲基-3H标记和35S标记的S-(8-叠氮腺苷)甲硫氨酸那样,用荧光增强剂浸渍丙烯酰胺凝胶以及将液体样品稀释到闪烁液中。