Identification and Distribution of Peptide:N-Glycanase (PNGase) in Mouse Organs

Kitajima, Ken; Suzuki, Tadashi; Kouchi, Zen; Inoue, Sadako; Inoue, Yousuke

doi:10.1006/abbi.1995.1309

Cited by 45 publications

(46 citation statements)

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“…The distal part of magnum portion of hen oviduct showed specific activity 5.5-7.5 times higher than those of the middle and proximal parts that showed the activities of 0.45-0.61 milliunit͞mg comparable to those detected in mammalian cells (15,17). Since in the distal portion massive amounts of egg white were secreted around eggs, high enzyme activity may be considered to be physiologically related to egg formation.…”

Section: Resultsmentioning

confidence: 76%

“…Recently, we have shown that soluble PNGase activities whose optimal activity lies at neutral pH occur ubiquitously in mammalian cultured cells and mouse organs (15)(16)(17). However, we have no definite information as to their endogenous substrates.…”

mentioning

confidence: 99%

“…The activity of PNGase was assayed using asialo-[ 14 C]fetGP I as a substrate as described (16,17). One unit of the PNGase activity was defined as the amount of enzyme releasing 1 mol of fetuin 14 C-labeled peptide I under the assay conditions at 25°C for 24 h.…”

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confidence: 99%

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Site-specific de- N -glycosylation of diglycosylated ovalbumin in hen oviduct by endogenous peptide: N -glycanase as a quality control system for newly synthesized proteins

Suzuki

Kitajima

Emori

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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Hen ovalbumin (OVA) is known to exist as a singly N-glycosylated form with a glycan chain on Asn-292 in egg white. Previous studies showed that di-N-glycosylated form of OVA [Di-OVA; CHO-Asn-292͞CHO-Asn-311 (CHO, N-glycan chain)], which has two N-glycan chains on Asn-292 and Asn-311, was expressed only transiently in hen oviduct. Di-OVA was not found in egg white, suggesting that this form cannot be secreted normally and may possibly be converted to mono-N-glycosylated OVA (CHO-Asn-292͞Asp-311) by the action of peptide:Nglycanase (PNGase) during synthesis and secretion. In this study, we have identified the putative PNGase activity in the homogenate of hen oviduct, purified 1,000-fold, and designated as PNGase HO. We examined the reactivity of Di-OVA to PNGase HO and found that this enzyme site-specifically cleaved off the glycan chain at Asn-311 to convert Di-OVA into the mono-N-glycosylated form (CHO-Asn-292͞Asp-311). In contrast, this enzyme was found not to act on the mono-Nglycosylated OVA (CHO-Asn-292͞Asn-311) found in egg white when it was tested as a substrate. The present findings support our view that de-N-glycosylation catalyzed by PNGase may be involved in quality control of newly synthesized proteins by converting its diglycosylated form into the mono-N-glycosylated form that can be secreted. However, the alternative possibility that de-N-glycosylation may trigger cytosolic degradation of the aberrantly glycosylated glycoprotein cannot be ruled out.

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Section: Resultsmentioning

confidence: 76%

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confidence: 99%

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Site-specific de- N -glycosylation of diglycosylated ovalbumin in hen oviduct by endogenous peptide: N -glycanase as a quality control system for newly synthesized proteins

Suzuki

Kitajima

Emori

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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“…5). Widespread expression has also been reported for plant and mammalian PNGases (Kitajima et al, 1995;Diepold et al, 2007), as might be expected for a housekeeping gene involved in glycoprotein catabolism. In the egg-laying system, promoter activity was detected consistently in all vulval cells from the late L3 stage to adulthood (Fig.…”

Section: Png-1 Is Widely Expressedmentioning

confidence: 72%

TheN-Glycanasepng-1Acts to Limit Axon Branching during Organ Formation in Caenorhabditis elegans

Habibi-Babadi

Su²,

Carvalho³

et al. 2010

J. Neurosci.

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Peptide:N-glycanases (PNGases) are cytoplasmic de-N-glycosylation enzymes that have been shown in cultured cells to facilitate the degradation of misfolded glycoproteins during endoplasmic reticulum-associated degradation and in the processing of major histocompatibility complex class I antigens for proper cell-surface presentation. The gene encoding PNGase activity was initially described in budding yeast (Png1p) and shown to be highly conserved from yeast to humans, but physiological roles in higher organisms have not been elucidated. Here we describe peripheral nervous system defects associated with the first loss-of-function mutations in an animal PNGase. Mutations in png-1, the Caenorhabditis elegans PNGase ortholog, result in an increase in axon branching during morphogenesis of the vulval egg-laying organ and egg-laying behavior changes. Neuronal defects include an increase in the branched morphology of the VC4 and VC5 egg-laying neurons as well as inappropriate branches from axons that run adjacent to the vulva but would normally remain unbranched. We show that png-1 is widely expressed and can act from both neurons and epithelial cells to restrict axon branching. A deletion allele of the DNA repair gene rad-23, orthologs of which are known to physically interact with PNGases in yeast and mammals, displays similar axon branching defects and genetic interactions with png-1. In summary, our analysis reveals a novel developmental role for a PNGase and Rad-23 in the regulation of neuronal branching during organ innervation.

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“…(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 as an enzyme but also as a carbohydrate recognition protein (i.e.…”

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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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Identification and Distribution of Peptide:N-Glycanase (PNGase) in Mouse Organs

Cited by 45 publications

References 0 publications

Site-specific de- N -glycosylation of diglycosylated ovalbumin in hen oviduct by endogenous peptide: N -glycanase as a quality control system for newly synthesized proteins

Site-specific de- N -glycosylation of diglycosylated ovalbumin in hen oviduct by endogenous peptide: N -glycanase as a quality control system for newly synthesized proteins

TheN-Glycanasepng-1Acts to Limit Axon Branching during Organ Formation in Caenorhabditis elegans

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

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