生化鉴定重组beta-glucosyltransferase和分析全球5-hydroxymethylcytosine独特的基因组。
文章的细节
-
引用
复制到剪贴板 -
郑Terragni J, Bitinaite J, Y,普拉丹
生化鉴定重组beta-glucosyltransferase和分析全球5-hydroxymethylcytosine独特的基因组。
生物化学,2012年2月7日,51(5):1009 - 19所示。doi: 10.1021 / bi2014739。Epub 2012年1月27日。
- PubMed ID
-
22229759 (在PubMed]
- 文摘
-
5-Hydroxymethylcytosine (5-hmC)是一种酶促氧化的产物5-methylcytosine (5-mC)。一千零一十一年的易位(春节)家族的酶催化转换5-mC 5-hmC。Phage-encoded葡糖基转移酶已知glucosylate 5-hmC,可以用来检测和分析5-hmC作为哺乳动物的表观基因组表观遗传标记。这里我们有进行了详细的生化特性和稳态动力学参数分析T4噬菌体beta-glucosyltransferase (beta-GT)。重组beta-GT glucosylates 5-hmC DNA nonprocessive的方式,和绑定5-hmC DNA或尿苷diphosphoglucose (UDP-glucose)基质是随机的,与二元配合物催化地能力。与beta-GT产物抑制的研究表明,UDP是一种竞争性抑制剂UDP-glucose和混合抑制剂对5-hmC DNA。同样,glucosylated-5-hmC (5-ghmC) DNA是一种竞争性抑制剂对5-hmC DNA和混合抑制剂对UDP-glucose。5-hmC DNA结合~ 10倍强beta-GT酶相比,其glucosylated产品。5-hmC目标序列的数量影响重组beta-GT营业额数字。此外,我们利用重组beta-GT估计全球5-hmC内容在不同的基因组dna。 Most of the genomic DNAs derived from vertebrate tissue and cell lines contained 5-hmC. DNA from mouse, human, and bovine brains displayed 0.5-0.9% of the total nucleotides as 5-hmC, which was higher compared to the levels found in other tissues. A comparison between cancer and healthy tissue genomes suggested a lower percentage of 5-hmC in cancer, which may reflect the global hypomethylation of 5-mC observed during oncogenesis.