Characterization of the peptide-N 4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) fromSilene alba cells

Lhernould, Sabine; Karamanos, Yannis; Lerouge, Patrice; Morvan, Henri

doi:10.1007/bf00731874

Cited by 23 publications

(11 citation statements)

References 30 publications

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“…in the case of autophagy (Lhernould et al, 1994). PNGase Se, that could be responsible for the formation of a portion of UNGs in white campion cells, was characterized (Lhernould et al, 1995).…”

Section: Plant Pngases: Possible Regulator Molecules In Cellular Procmentioning

confidence: 99%

Occurrence and Potential Functions ofN–Glycanases

Suzuki¹,

Kitajima²,

Inoue³

et al. 1996

Glycosciences

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Section: Plant Pngases: Possible Regulator Molecules In Cellular Procmentioning

confidence: 99%

Occurrence and Potential Functions ofN–Glycanases

Suzuki¹,

Kitajima²,

Inoue³

et al. 1996

Glycosciences

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“…[25][26][27][28][29][30]. Unconjugated N-glycans (29) were isolated from the extracellular medium of a plant-cell suspension of white campion, Silene alba (25,26) as well as from the tomato fruit pericarp (Ref.…”

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confidence: 99%

Developmentally Regulated Expression of a Peptide:N-Glycanase during Germination of Rice Seeds (Oryza sativa) and Its Purification and Characterization

Chang

Kuo

Khoo

et al. 2000

Journal of Biological Chemistry

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Peptide:N-glycanase (PNGase; EC 3.5.1.52) activity was detected in dormant rice seeds (Oryza sativa) and the imbibed rice grains. Time-course studies revealed that the enzyme activity remained almost constant until about 30 h after imbibition in both of endosperm-and embryo tissue-containing areas, and started to increase only in growing germ part, reached a peak at about 3-day stage, followed by a gradual decrease concomitant with a sharp increase in the coleoptile. The specific activity increased about 6-fold at about 3-day stage. PNGase was purified to electrophoretic homogeneity from the extracts of germinated rice seeds at 24 h, and the apparent molecular weight of the purified enzyme, estimated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), was about 80,000. The purified enzyme was designated PNGase Os to denote its origin. The N-terminal sequence of the 10 residues was determined to be SYN-VASVAGL. The purified PNGase Os in SDS-PAGE appeared as a rather broad band, consistent with the presence of multiple glycoforms as indicated by chromatographic behavior on a Sephadex G-75 column. PNGase expressed in coleoptile under anoxia condition was also purified, and both of the purified enzymes were found to exhibit very similar, if not identical, electrophoretic mobility in SDS-PAGE. PNGase Os exhibited a broad pH-activity profile with an optimum of 4 -5 and, interestingly, was significantly inactivated by K ؉ and Na ؉ at near the physiological concentration, 100 mM. These results are discussed in relation to other work.-(N-acetyl-␤-Dglucosaminyl) asparagine amidase, EC 3.5.1.52) had only been known to occur in some plant seeds (1-5) and bacteria (6) and used as a useful reagent in a number of studies of structure and function of glycoproteins having N-linked glycan chains until we demonstrated for the first time its occurrence in the early embryos of Medaka fish, Oryzias latipes in 1991 (7). Following this discovery, we began to focus our interest on the physiological significance of this enzyme in living organisms because little attention had been paid to it. In view of the unique structural change in a given functional protein by converting the glycosylated asparagine residue to the aspartic acid residue upon de-N-glycosylation catalyzed by PNGase as the posttranslational remodification of protein, we anticipated that N-glycosylation of proteins by oligosaccharyltransferase and their de-N-glycosylation by PNGase constitute a basic biological mechanism of functioning within cells (8, 9). For understanding the biological meaning of the occurrence of PNGase in eukaryotes, we have been involved in a series of studies on animal-derived PNGases (10 -15) and reported the following. (a) PNGase activities were shown to occur in mammalianderived cell lines including human origin (10). (b) PNGase activities were detected ubiquitously in various organs and tissues of mouse (13). (c) PNGase was purified to homogeneity from the confluent stage of C3H mouse fibroblast L-929 cells and characterized to serve not only a...

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“…Such a conformational change could regulate the interaction between Con A and the jackbean N-glycanase. Preliminary analyses of N-glycanase activities from Raphanus sati us [28] and Silene alba [44] have shown they also bind to immobilized Con A matrices, although, as yet, characterization of the enzymes has not been carried out.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of N-glycanase, a concanavalin A binding protein from jackbean (Canavalia ensiformis)

Sheldon

Keen

Bowles

1998

Biochemical Journal

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Removal of the N-glycan from the concanavalin A (Con A) glycoprotein precursor is a key step in its conversion into an active lectin. N-Glycanase (EC 3.5.1.52), the enzyme from jackbean catalysing this process, has been purified to homogeneity as judged by native PAGE. One of the purification steps is binding of the enzymic activity to Con A-Sepharose and its elution by methyl α-mannoside. On SDS/PAGE the principal components were found to be 78 kDa, 74 kDa, 54 kDa, 32 kDa and 30 kDa polypeptides. These did not react with Con A on an affinity blot. Cleveland mapping indicated that some of these polypeptides had related primary structures. The enzyme has a broad pH optimum in the region of 5.0.

show abstract

Characterization of the peptide-N 4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) fromSilene alba cells

Cited by 23 publications

References 30 publications

Occurrence and Potential Functions ofN–Glycanases

Occurrence and Potential Functions ofN–Glycanases

Developmentally Regulated Expression of a Peptide:N-Glycanase during Germination of Rice Seeds (Oryza sativa) and Its Purification and Characterization

Purification and characterization of N-glycanase, a concanavalin A binding protein from jackbean (Canavalia ensiformis)

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