Wolfes H, Fliess A, Winkler F, Pingoud A
Eur J Biochem. 1986 Sep 1;159(2):267-73. doi: 10.1111/j.1432-1033.1986.tb09863.x.
We have synthesized several self-complementary oligodeoxynucleotides which contain bromodeoxyuridine in various positions within and outside of the recognition sequence for the EcoRI and EcoRV restriction endonucleases. These oligodeoxynucleotides are cleaved in the presence of Mg2+ by their respective enzyme. Upon irradiation by long-wavelength ultraviolet light and in the absence of Mg2+ they are cross-linked in low yield to their enzymes, forming 1:1 and 1:2 (oligodeoxynucleotide:enzyme subunit) adducts. Cross-linking occurs with both specific and non-specific complexes. With EcoRI the site of cross-linking was determined to be at or close to Met-137, i.e. in a region of the molecule implicated by other studies from our laboratory [Scholtissek et al. (1986) J. Biol. Chem. 261, 2228-2234] in the binding and cleavage of the substrate.
我们合成了几种自身互补的寡脱氧核苷酸，这些寡核苷酸在EcoRI和EcoRV限制性内切酶识别序列的内部和外部的不同位置含有溴脱氧尿苷。这些寡脱氧核苷酸在Mg2+存在下会被各自的酶切割。在长波长紫外光照射下且不存在Mg2+时，它们会以低产率与各自的酶发生交联，形成1:1和1:2（寡脱氧核苷酸:酶亚基）加合物。交联发生在特异性和非特异性复合物中。对于EcoRI，交联位点被确定在Met-137处或其附近，即在我们实验室其他研究[Scholtissek等人（1986年）《生物化学杂志》261卷，2228 - 2234页]所涉及的分子区域，该区域与底物的结合和切割有关。
