PEPTIDE-N4-(N-ACETYL-BETA-GLUCOSAMINYL)ASPARAGINE AMIDASE-F CANNOT RELEASE GLYCANS WITH FUCOSE ATTACHED ALPHA-1-]3 TO THE ASPARAGINE-LINKED N-ACETYLGLUCOSAMINE RESIDUE
PEPTIDE-N4-(N-ACETYL-BETA-GLUCOSAMINYL)ASPARAGINE AMIDASE-F CANNOT RELEASE GLYCANS WITH FUCOSE ATTACHED ALPHA-1-]3 TO THE ASPARAGINE-LINKED N-ACETYLGLUCOSAMINE RESIDUE
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DOI：
10.1111/j.1432-1033.1991.tb16166.x
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发表时间：
1991-08-01
期刊：
影响因子：
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通讯作者：
MARZ, L
中科院分区：
文献类型：
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作者：
TRETTER, V;ALTMANN, F;MARZ, L
比较了脑膜炎黄杆菌肽- n4 -（n -乙酰- β -氨基葡萄糖）天冬酰胺酰胺酶F （PNGase F）和甜杏仁肽- PNGase A对植物n -糖肽和n -糖蛋白去糖基化的能力。以菠萝蛋白酶糖肽和辣根过氧化物酶- c糖蛋白为底物，其中木糖连接β -1 -> - 2与β -甘露糖相连，α -1 -> - 3与最内层的n -乙酰氨基葡萄糖相连。与PNGase A相反，来自脑膜炎脓毒杆菌的酶即使在比常用标准底物完全去糖基化所需的浓度高100倍的浓度下也不作用于这些底物。通过温和的酸水解去除植物糖肽和糖蛋白中的α -1 -> 3-linked focal后，它们在中等浓度的酶下很容易被PNGase F降解。因此，我们得出结论，内部n -乙酰氨基葡萄糖的α -1 - b>3集中化阻碍了PNGase F的酶促作用。了解PNGase F去糖基化潜力的这一限制可能会使它从一个陷阱变成一个有用的实验工具。
The ability of peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F (PNGase F) from Flavobacterium meningosepticum and PNGase A from sweet almonds to deglycosylate N-glycopeptides and N-glycoproteins from plants was compared. Bromelain glycopeptide and horseradish peroxidase-C glycoprotein, which contain xylose linked beta-1 --> 2 to beta-mannose and fucose linked alpha-1 --> 3 to the innermost N-acetylglucosamine, were used as substrates.In contrast to PNGase A, the enzyme from F. meningosepticum did not act upon these substrates even at concentrations 100-fold higher than required for complete deglycosylation of commonly used standard substrates. After removal of alpha-1 --> 3-linked fucose from the plant glycopeptide and glycoprotein by mild acid hydrolysis, they were readily degraded by PNGase F at moderate enzyme concentrations.Hence we conclude that alpha-1 --> 3 fucosylation of the inner N-acetylglucosamine impedes the enzymatic action of PNGase F. Knowledge of this limitation of the deglycosylation potential of PNGase F may turn it from a pitfall into a useful experimental tool.