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METHODS AND KITS FOR DETECTING O-GlcNAc SITES USING B3GALNT2 AND OGT
METHODS AND KITS FOR DETECTING O-GlcNAc SITES USING B3GALNT2 AND OGT
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机译:B3GALNT2和OGT检测 O I> -GlcNAc位点的方法和试剂盒
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摘要
In an in vitro method of determining the modification degree of O-GlcNAc in a biological sample, a first sample is treated with B3GALNT2 to incorporate a GalNAz into closed O-GlcNAc sites in the sample, labeled using click chemistry to form labeled closed O-GlcNAc sites, and the labeled closed O-GlcNAc sites in the first sample are detected (corresponding to the number of closed O-GlcNAc sites in the biological sample). A second, duplicate sample is treated with OGT to O-GlcNAcylate open O-GlcNAc sites in the sample to convert the open O-GlcNAc sites to closed O-GlcNAc sites, and treated with B3GALNT2 to incorporate GalNAz into the closed O-GlcNAc sites. The second sample is labeled using click chemistry to form labeled closed O-GlcNAc sites (corresponding to the total number of O-GlcNAc sites in the biological sample). The number of closed O-GlcNAc sites and total O-GlcNAc sites in the biological sample are compared to determine the modification degree of O-GlcNAc.
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机译:在确定生物样品中 O I> -GlcNAc修饰度的体外方法中,第一个样品用B3GALNT2处理,将GalNAz掺入封闭的 O I> -GlcNAc位点在样品中,使用点击化学方法标记以形成标记的封闭 O I> -GlcNAc位点,并且在第一个样本中检测到标记的封闭 O I> -GlcNAc位点(对应于数量生物样品中封闭的 O I> -GlcNAc位点的序列)。用OGT将第二个重复的样品处理为样品中的 O I> -GlcNAcylate开放 O I> -GlcNAc位点,以转换开放的 O I> -GlcNAc位点到封闭的 O I> -GlcNAc位点,并用B3GALNT2处理,将GalNAz掺入封闭的 O I> -GlcNAc位点。使用点击化学法标记第二个样品,形成标记的封闭 O I> -GlcNAc位点(对应于生物样品中 O I> -GlcNAc位点的总数)。比较生物样品中封闭的 O I> -GlcNAc位点的数量和总 O I> -GlcNAc位点的数量,以确定 O I> -GlcNAc的修饰度。
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