一种用于检测作用于天冬酰胺连接聚糖中二-N-乙酰壳二糖部分的酶的荧光高效液相色谱测定法。
A fluorescence high-performance liquid chromatography assay for enzymes acting on the di-N-acetylchitobiosyl part of asparagine-linked glycans.
作者信息
Bourgerie S, Berger S, Strecker G, Julien R, Karamanos Y
机构信息
Institut de Biotechnologie, Université de Limoges, France.
出版信息
J Biochem Biophys Methods. 1994 Jun;28(4):283-93. doi: 10.1016/0165-022x(94)90004-3.
The glycoasparagine, Man7GlcNAc2Asn ('Man7') was labelled with resorufin and used as a specific substrate for the detection and quantification of endo-beta-N-acetyl glucosaminidases (Endos) acting on the di-N-acetylchitobiosyl part of asparagine-linked glycans. Peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidases (PNGases) cannot transform this substrate but they can be detected by the procedure described earlier using the resorufin-labelled N-glycopeptide [Glycoconjugate J., 9 (1992) 162-167]. These two substrates can be used in a simple, reproducible and very sensitive fluorescence HPLC assay in order to monitor Endo and PNGase activities during isolation and purification processes, or studies of the evolution of such activities during cultivation of the producing cells.
糖天冬酰胺，Man7GlcNAc2Asn（“Man7”）用试卤灵标记，并用作检测和定量作用于天冬酰胺连接聚糖的二 - N - 乙酰壳二糖部分的内切β - N - 乙酰葡糖胺糖苷酶（Endos）的特异性底物。肽 - N4 - （N - 乙酰 - β - 葡糖胺基）天冬酰胺酰胺酶（PNGases）不能转化该底物，但可以通过先前使用试卤灵标记的N - 糖肽所述的方法进行检测[《糖缀合物杂志》，9（1992）162 - 167]。这两种底物可用于简单、可重复且非常灵敏的荧光HPLC分析，以便在分离和纯化过程中监测Endo和PNGase活性，或研究生产细胞培养过程中此类活性的演变。
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