A FLUORESCENCE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ASSAY FOR ENZYMES ACTING ON THE DI-N-ACETYLCHITOBIOSYL PART OF ASPARAGINE-LINKED GLYCANS

被引：9

作者：

BOURGERIE, S

BERGER, S

STRECKER, G

JULIEN, R

KARAMANOS, Y

机构：

[1] UNIV LIMOGES,INST BIOTECHNOL,F-87060 LIMOGES,FRANCE

[2] UNIV SCI & TECH LILLE FLANDRES ARTOIS,CHIM BIOL LAB,F-59655 VILLENEUVE DASCQ,FRANCE

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1994年 / 28卷 / 04期

关键词：

ENDOGLYCOSIDASE ASSAY; ENDO; PNGASE; FLUORESCENCE-HPLC;

D O I：

10.1016/0165-022X(94)90004-3

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The glycoasparagine, Man(7)GlcNAc(2)Asn ('Man(7)') 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-N-4-(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.

引用

页码：283 / 293

页数：11

共 9 条

[1] USE OF RESORUFIN-LABELED N-GLYCOPEPTIDE IN A HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ASSAY TO MONITOR ENDOGLYCOSIDASE ACTIVITIES DURING CULTIVATION OF FLAVOBACTERIUM-MENINGOSEPTICUM [J].