Information on EC 3.5.1.52 - peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
3.5.1.52
-
RECOMMENDED NAME
GeneOntology No.
peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Hydrolysis of an N4-(acetyl-beta-D-glucosaminyl)asparagine residue in which the glucosamine residue may be further glycosylated, to yield a (substituted) N-acetyl-beta-D-glucosaminylamine and a peptide containing an aspartate residue
show the reaction diagram
-
-
-
-
Hydrolysis of an N4-(acetyl-beta-D-glucosaminyl)asparagine residue in which the glucosamine residue may be further glycosylated, to yield a (substituted) N-acetyl-beta-D-glucosaminylamine and a peptide containing an aspartate residue
show the reaction diagram
Asn-Asn(oligosaccharide)-Glu-Ser-Ser + H2O = Asn-Asp-Glu-Ser-Ser + 1-amino-N-acetylglucosamine-oligosaccharide, step 1, catalysed by the enzyme, 1-amino-N-acetylglucosamine-oligosaccharide + H2O = N-acetylglucosamine-oligosaccharide + NH3, step 2
Prunus amygdalus var. dulcis
-
Hydrolysis of an N4-(acetyl-beta-D-glucosaminyl)asparagine residue in which the glucosamine residue may be further glycosylated, to yield a (substituted) N-acetyl-beta-D-glucosaminylamine and a peptide containing an aspartate residue
show the reaction diagram
mechanism and catalytic center
-
Hydrolysis of an N4-(acetyl-beta-D-glucosaminyl)asparagine residue in which the glucosamine residue may be further glycosylated, to yield a (substituted) N-acetyl-beta-D-glucosaminylamine and a peptide containing an aspartate residue
show the reaction diagram
the catalytic triad is formed by Cys191, His218, and Asp235
-
Hydrolysis of an N4-(acetyl-beta-D-glucosaminyl)asparagine residue in which the glucosamine residue may be further glycosylated, to yield a (substituted) N-acetyl-beta-D-glucosaminylamine and a peptide containing an aspartate residue
show the reaction diagram
active site structure and catalytic mechanism
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
Q5XI55
-
hydrolysis
-
-
hydrolysis
Q02890
-
hydrolysis of carboxylic acid amide
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
N-linked-glycopeptide-(N-acetyl-beta-D-glucosaminyl)-L-asparagine amidohydrolase
Does not act on (GlcNAc)Asn, because it requires the presence of more than two amino-acid residues in the substrate [cf. EC 3.5.1.26, N4-(beta-N-acetylglucosaminyl)-L-asparaginase]. The plant enzyme was previously erroneously listed as EC 3.2.2.18.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acid PNGase M
-
-
-
-
acidic peptide:N-glycanase
D0QU16
-
acidic PNGase
D0QU16
-
glycoamidase
D0QU16
-
glycopeptidase
-
-
-
-
glycopeptide N-glycosidase
-
-
-
-
jackbean glycopeptidase
-
-
-
-
L-929 PNGase
-
-
-
-
MPng1
-
-
N-glycanase
-
-
-
-
N-oligosaccharide glycopeptidase
-
-
-
-
neutral PNGase M
-
-
-
-
peptide N-glycanase
-
-
peptide N-glycanase
-
-
peptide N-glycanase
-
-
peptide N-glycosidase F
-
-
peptide N4(N-acetyl-glucosaminyl)asparagine amidase
-
-
peptide-N-(N-acetyl-beta-glucosaminyl) asparagine amidase F
-
-
peptide-N-glycanase
-
-
peptide-N-glycosidase F
-
-
peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine amidase F
-
-
-
-
peptide: N-glycanase
-
-
peptide: N-glycanase
-
-
peptide: N-glycanase
-
-
peptide:N-glycanase
-
-
peptide:N-glycanase
-
-
peptide:N-glycanase
Q9TW67
-
peptide:N-glycanase
-
-
peptide:N-glycanase
-
-
peptide:N-glycanase
-
-
peptide:N-glycanase
Q9JI78
-
peptide:N-glycanase
-
-
peptide:N-glycanase
Q02890
-
peptide:N-glycanase
-
-
peptidyl N-glycanase F
-
-
PNG1
-
-
PNG1
-
wild type PNG1 has substitutions in essential catalytic amino acids, and its deglycosylation activity is lost
PNGase
Q9JI78
-
PNGase A
-
-
-
-
PNGase At
-
-
-
-
PNGase F
-
-
-
-
PNGase F
-
-
PNGase F
Q5XI55
-
PNGase J
-
-
-
-
PNGase Os
-
-
-
-
PNGase Se
-
-
-
-
PNGaseF
-
-
yeast peptide: N-glycanase
-
-
CAS REGISTRY NUMBER
COMMENTARY
83534-39-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
ecotype Columbia 0
-
-
Manually annotated by BRENDA team
expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
gene png1
-
-
Manually annotated by BRENDA team
rice
-
-
Manually annotated by BRENDA team
Prunus amygdalus var. dulcis
almond
-
-
Manually annotated by BRENDA team
several strains, overview
-
-
Manually annotated by BRENDA team
cultivar Momotaro
UniProt
Manually annotated by BRENDA team
Mill. cv. Dombito
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
PNG1 deletion results in strong polarity defects
malfunction
-
mutations in png-1 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
malfunction
-
homozygous deletion mutants show developmental defect
physiological function
-
the peptide:N-glycanase ortholog PNG1 is essential for cell polarity despite its lack of enzymatic activity, PNG1 is involved in cell wall integrity
physiological function
-
png-1 can act from both neurons and epithelial cells to restrict axon branching. PNGase and Rad-23 regulate neuronal branching during organ innervation
physiological function
-
the complex formed between peptide:N-glycanase and Rad23p exhibits enhanced deglycosylation activity, which suggests an important role for this enzyme in the misfolded glycoprotein degradation pathway in vivo
physiological function
-
the enzyme is a deglycosylating enzyme involved in the endoplasmic reticulum-associated degradation process
physiological function
-
the N-terminal domain of PNGase (PUB) serves as a possible activator of HR23 in endoplasmic reticulum-associated degradation mechanisms
physiological function
D0QU16, -
the enzyme is involved in a different de-N-glycosylation mechanism associated with plant growth and development
physiological function
-
the enzyme plays a critical role during larval development and metamorphosis
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetylcholine receptor + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-dansyl derivative fetuin pentaglycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
alpha1-antitrypsin Pi Z + H2O
deglycosylated alpha1-antitrypsin Pi Z
show the reaction diagram
-
-
-
?
Antithrombin III + H2O
?
show the reaction diagram
-
significant amount of only partly de-N-glycosylated protein is detected even after more than 18 h incubation time with PNGase F
-
-
?
asialoglycopeptide I + H2O
?
show the reaction diagram
-
fetuin-derived radio-labeled substrate, deglycosylation
-
-
?
ATPase p97 + H2O
?
show the reaction diagram
Q9JI78
a cofactor-binding motif of p97 contained within the last 10 amino acid residues of the C terminus is both necessary and sufficient to mediate interactions of p97 with PNGase. Phosphorylation of p97s highly conserved penultimate tyrosine residue, which is the main phosphorylation site during T cell receptor stimulation, completely blocks binding of either PNGase or Ufd3 to p97. This observation suggests that phosphorylation of this residue modulates endoplasmic reticulum-associated protein degradation activity by discharging substrate-processing cofactors, a cofactor-binding motif of p97 contained within the last 10 amino acid residues of the C terminus is both necessary and sufficient to mediate interactions of p97 with PNGase
-
-
?
bence-jones wh lambda glycopeptide + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11
-
-
?
beta-aspartylglycosylamine + H2O
aspartic acid + NH3 + N-acetylglucosamine
show the reaction diagram
-
1-L-beta-aspartamido-2-acetamido-1,2-dideoxy-beta-D-glucose
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
Leu-Asn(Man3Gal3GlcNAc5)-Asp-Ser-Arg
-
-
-
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
Leu-Asn(Man3Gal3GlcNAc5)-Asp-Ser-Arg
-
-
?
bovine fetuin glycopeptide + H2O
?
show the reaction diagram
-
Leu-Ala-Asn-(oligosaccharide)-AeCys-Ser
-
-
?
class I MHC HC + H2O
?
show the reaction diagram
-
the enzyme is involved proteasomal degradation of glycosylated type I membrane protein class I MHC heavy chain, dislocated from ER to cytosol by cytomegalovirus-encoded glycoprotein US2 protein, overview, the enzyme deglycosylates dislocated membrane protein class I MHC heavy chain
-
-
?
coagulation factor IX + H2O
?
show the reaction diagram
-
almost quantitative de-N-glycosylation with PNGase F is observed after a rather short time
-
-
?
concanavalin A-precursor + H2O
?
show the reaction diagram
-
deglycosylation leads to conversion into an active lectin
-
-
?
Cys-Gly-Leu-Val-Pro-Val-Leu-Ala-Glu-Asn-Tyr-Asn(Man3Gal2GlcNAc4NeuAc2)-Lys + H2O
?
show the reaction diagram
D0QU16, -
-
-
-
?
dedansylated hen ovomucoid glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
dedansylated hen ovomucoid glycopeptide + H2O
?
show the reaction diagram
-
DGP
-
-
?
denatured ribonuclease B + H2O
?
show the reaction diagram
-
-
-
-
?
denatured RNase B + H2O
?
show the reaction diagram
-
SpPNGase cleaves N-glycan from denatured RNase B
-
-
?
denatured RNaseB + H2O
deglycosylated RNaseB
show the reaction diagram
-
-
-
?
fetuin glycopeptide II + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11
-
-
?
fetuin-derived asialoglycopeptide I + H2O
?
show the reaction diagram
-
([14C]-CH3)2Leu-Asn(GlcNAc5-Man3Gal3)-Asp-Ser-Arg
-
-
?
Gal2GlcNAc2Man3GlcNAc2-Asn-peptide + H2O
?
show the reaction diagram
-
-
-
-
?
GlcNAc-Asn-peptide + H2O
?
show the reaction diagram
-
-
-
-
?
glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
glycopeptide + H2O
?
show the reaction diagram
Prunus amygdalus var. dulcis
-
with 3-11 amino acid residues
-
-
?
glycoprotein US11 + H2O
?
show the reaction diagram
-
the enzyme deglycolyzes HCMV glycoprotein US11
-
-
?
glycoprotein US2 + H2O
?
show the reaction diagram
-
the enzyme deglycolyzes HCMV glycoprotein US2
-
-
?
HR23-ubiquitin-like domain + H2O
?
show the reaction diagram
-
the N-terminal domain of PNGase (PUB) interacts with HR23-ubiquitin-like domain and ubiquitin chains
-
-
?
human asialotransferrin + H2O
?
show the reaction diagram
-
-
-
-
?
human IgG + H2O
?
show the reaction diagram
-
-
-
-
?
human transferrin glycopeptide + H2O
?
show the reaction diagram
D0QU16, -
-
-
-
?
L-hyosophorin + H2O
?
show the reaction diagram
-
neutral and acid PNGase M
-
-
?
laccase + H2O
?
show the reaction diagram
-
-
-
-
?
Leu-Asn(GlcNAc5Man3Gal3)-Asp-Ser-Arg + H2O
?
show the reaction diagram
Q02890
fetuin-derived asialoglycopeptide I, 14C, substrate activity assay
-
-
?
Man5GlcNAc2-Asn-peptide + H2O
?
show the reaction diagram
-
-
-
-
?
mu opioid receptor + H2O
?
show the reaction diagram
-
the enzyme removes all N-linked glycans
-
-
?
N-glycoprotein + H2O
?
show the reaction diagram
-
PNGase is involved in the release of N-glycans from N-glycoproteins
-
-
?
Oryzias latipes glycophosphoprotein MU-1 + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11, yolk-absorptive stage
-
-
?
Oryzias latipes glycophosphoprotein MU-2 + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11, yolk-absorptive stage
-
-
?
ovalbumin + H2O
?
show the reaction diagram
-
deglycosylation of the heat-denatured protein
-
-
?
ovalbumin glycopeptide + H2O
aspartic acid + NH3 + N-acetylglucosamine
show the reaction diagram
-
Glu-Glu-Lys-Tyr-Asn(oligosaccharide)-Leu-Thr-Ser-Val
-
?
ovalbumin glycopeptide + H2O
aspartic acid + NH3 + N-acetylglucosamine
show the reaction diagram
-
Glu-Glu-Lys-Tyr-Asn(oligosaccharide)-Leu-Thr-Ser-Val
-
-
?
ovalbumin glycopeptide + H2O
aspartic acid + NH3 + N-acetylglucosamine
show the reaction diagram
Prunus amygdalus var. dulcis
-
Glu-Glu-Lys-Tyr-Asn(oligosaccharide)-Leu-Thr-Ser-Val
-
?
ovalbumin glycopeptide + H2O
aspartic acid + NH3 + N-acetylglucosamine
show the reaction diagram
-
Glu-Glu-Lys-Tyr-Asn(oligosaccharide)-Leu-Thr-Ser-Val
-
-
?
ovotransferrin glycopeptide + H2O
?
show the reaction diagram
Prunus amygdalus var. dulcis
-
Gly-Leu-Ile-His-Asn(oligosaccharide)-Arg
-
-
?
PHA glycopeptide + H2O
?
show the reaction diagram
-
-
-
-
?
pineapple stem bromelain glycopeptide + H2O
?
show the reaction diagram
-
Asn-Asn-(oligosaccharide)-Glu-Ser-Ser
-
-
?
pineapple stem bromelain glycopeptide + H2O
?
show the reaction diagram
-
Asn-Asn-(oligosaccharide)-Glu-Ser-Ser
-
-
?
pineapple stem bromelain glycopeptide + H2O
?
show the reaction diagram
Prunus amygdalus var. dulcis
-
Asn-Asn-(oligosaccharide)-Glu-Ser-Ser
-
-
?
pineapple stem bromelain glycopeptide + H2O
?
show the reaction diagram
-
Asn-Asn-(oligosaccharide)-Glu-Ser-Ser
-
-
?
porcine fibrinogen + H2O
?
show the reaction diagram
-
-
-
-
?
proton-coupled folate transporter + H2O
?
show the reaction diagram
-
-
-
-
?
receptor for advanced glycation end-products isoform H-300 + H2O
?
show the reaction diagram
-
-
-
-
?
receptor for advanced glycation end-products isoform N-16 + H2O
?
show the reaction diagram
-
-
-
-
?
ribonuclease B + H2O
?
show the reaction diagram
-
no activity with bovine pancreatic ribonuclease B with oligomannose-type N-glycans at Asn-34. Native RNase BS, generated by subtilisin digestion of native RNase B, which comprises amino acid residues 21-124 of RNase B, is sensitive to PNGase F digestion. The same holds for carboxymethylated RNase. The structural basis of the difference in sensitivity for PNGase F in the de-N-glycosylation of the native bovine pancreatic ribonucleases B and BS is determined
-
-
?
ribonuclease B + H2O
?
show the reaction diagram
-
When ribonuclease B is denatured at 60-65C or by 40-60 mM dithiothreitol, its deglycosylation by Png1p is most prominent
-
-
?
ricin A + H2O
?
show the reaction diagram
-
deglycosylation of ricin A, the enzyme acts in complex with protein Rad23, interaction and complex formation analysis, overview
-
-
?
RNAse + H2O
?
show the reaction diagram
-, Q9TW67
removes high mannose-type N-glycan from denatured RNase B, but not from native RNase B
-
-
?
RNase B + H2O
?
show the reaction diagram
-
-
-
-
?
RNase B + H2O
?
show the reaction diagram
-
-
-
-
?
RNase B + H2O
?
show the reaction diagram
-
-
-
-
?
RNase B + H2O
?
show the reaction diagram
-
the enzyme deglycolyzes RNaseB
-
-
?
RTADELTA protein + H2O
deglycosylated RTADELTA protein + ?
show the reaction diagram
-
RTADELTA is a non-toxic mutant of ricin A-chain
-
-
?
RTADELTA-transmembrane-Leu2 protein + H2O
deglycosylated RTADELTA-transmembrane-Leu2 protein + ?
show the reaction diagram
-
-
-
-
?
sialylglycopeptide + H2O
?
show the reaction diagram
-, Q9TW67
asialo-, agalactosyl-, trimannosyl- and monomanosyl-sialylglycopeptides are hydrolyzed
-
-
?
taka-amylase A + H2O
?
show the reaction diagram
Prunus amygdalus var. dulcis
-
1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1
-
-
?
transferrin + H2O
?
show the reaction diagram
Prunus amygdalus var. dulcis
-
desialylated, human
-
-
?
truncated RNaseB + H2O
deglycosylated RNaseB
show the reaction diagram
-
-
-
?
tyrosinase + H2O
?
show the reaction diagram
-
the enzyme is required for processing of a class I-restricted epitope from tyrosinase together with the cooperative action of endoplasmic reticulum aminopeptidase 1 and cytosolic protease, overview, deglycosylation of tyrosinase at Asn371 and the Tyr369 epitope, reduced activity with tyrosinase peptides that are mutated in the glycosylation consensus sequence to give T373V mutants
-
-
?
XylMan3FucGlcNAc2-Asn-peptide from horse raddish peroxidase + H2O
?
show the reaction diagram
-
-
-
-
?
yeast carboxypeptidase + H2O
?
show the reaction diagram
-
deglycosylation of heat-denatured protein
-
-
?
melanopsin + H2O
?
show the reaction diagram
Q5XI55
-
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
-
-
-
-
additional information
?
-
-
-
-
-
-
additional information
?
-
-
-
-
-
-
additional information
?
-
Prunus amygdalus var. dulcis
-
-
-
-
-
additional information
?
-
-
ability of enzyme to distinguish between native and misfolded substrates
-
?
additional information
?
-
-
overview on in vivo glycoprotein substrates, involved in proteasomal degradation of misfolded glycoproteins
-
?
additional information
?
-
-
the cytosolic enzyme deglycosylates misfolded glycoproteins prior to the endoplasmic reticulum-associated protein degradation
-
-
-
additional information
?
-
-
the enzyme deglycosylates misfolded glycoproteins, the enzyme is a mediator for p97 functions, the PUB domain functions as a p97 binding module in human enzyme, p97 is an AAA ATPase with an ubiquitin-selective molecular machine involved in multiple cellular processes including protein degradation through the ubiquitin-proteasome system
-
-
-
additional information
?
-
-
the enzyme is responsible for deglycosylation of N-linked glycoproteins dislocated from endoplasmic reticulum to cytosol
-
-
-
additional information
?
-
-
the enzyme mediates the binding of the cytoplasmic proteins p97 and HR23B to the proteasome through formation of a ternary complex, p97 binds the autocrine motility factor receptor AMFR, modeling of interaction of the endoplasmic reticulum with the proteasome, overview
-
-
-
additional information
?
-
-
the enzyme removes N-linked oligosaccharides from misfolded glycoproteins as part of endoplasmic reticulum-associated degradation pathway involving a complex formation with proteins HR23B, cytosolic protein Y33K, p97, and autocrine motility factor receptor AMFR, the AAA ATPase p97 links peptide N-glycanase to the endoplasmic reticulum-associated E3 ligase AMFR, the N-terminus of PNGase interacts with the C-terminal tail of AMFR, complex formation model, overview
-
-
-
additional information
?
-
-
the enzyme removes N-linked oligosaccharides from misfolded glycoproteins as part of endoplasmic reticulum-associated degradation pathway involving a tight complex-formation with protein HR23, HR23 is also involved in DNA repair, co-evolution of the endoplasmic reticulum-associated degradation and DNA repair pathways, overview
-
-
-
additional information
?
-
-
PNGase F is responsible for deglycosylation of misfolded proteins, the reverse reaction, glycosylation of peptides RKDVY and EILDVPST by PNGase F, is also possible in vitro, determination of glycosylation sites by MALDI-TOF mass spectrometry, overview, in addition non-enzymatic glycosylation of sugars, which are more than two units long, occurs
-
-
-
additional information
?
-
-
the deglycosylating enzyme catalyzes the hydrolysis of the beta-aspartylglycosylamine bond of asparagine-linked glycopeptides and glycoproteins, the misfolding or denaturation of glycoproteins is a prerequisite for enzyme activity
-
-
-
additional information
?
-
-
the enzyme deglycosylates misfolded proteins in the cytosol and shows multiple modes of interaction with the proteasome, overview
-
-
-
additional information
?
-
-
the enzyme hydrolyzes the beta-aspartylglycosylamine bond of asparagine-linked glycopeptides and glycoproteins, releasing an intact oligosaccharide and generating an aspartic acid residue at the cleavage site, the N-terminus of the enzyme interacts with the DNA repair protein Rad23 detected by gel filtration and surface plasmon resonance, quantitation of complex formation
-
-
-
additional information
?
-
-
the enzyme is a glycoaminidase cleaving the link between asparagine and N-acetylglucosamines
-
-
-
additional information
?
-
-
SpPNGase deglycosylates the misfolded glycoproteins
-
-
-
additional information
?
-
-
the AtPNG1 gene encodes a bona fide peptide:N-glycanase that contributes to ERAD related protein quality control in plants
-
-
-
additional information
?
-
-
PNG-1, the cytoplasmic PNGase orthologue, exhibits dual enzyme functions, not only as peptide:N-glycanase but also as an oxidoreductase(thioredoxin)
-
-
-
additional information
?
-
Q02890
catalyses the de-glycosylation of unfolded glycoproteins
-
-
-
additional information
?
-
-
peptide:N-glycanase catalyzes the detachment of N-linked glycan chains from glycopeptides or glycoproteins by hydrolyzing the beta-aspartylglucosaminyl bond. PNGase can not deglycosylate correctly folded native glycoproteins, but catalyzes the deglycosylation of misfolded glycoproteins
-
-
-
additional information
?
-
-
peptide:N-glycanase catalyzes the detachment of N-linked glycan chains from glycopeptides or glycoproteins by hydrolyzing the beta-aspartylglucosaminyl bond. PNGase can not deglycosylate correctly folded native glycoproteins, but catalyzes the deglycosylation of misfolded glycoproteins. The complex formed between peptide:N-glycanase and Rad23p exhibits enhanced deglycosylation activity
-
-
-
additional information
?
-
D0QU16, -
the enzyme hydrolyses the beta-aspartyl-glycosylamine bond of N-linked glycoproteins/glycopeptides and releases free N-glycans
-
-
-
additional information
?
-
-
PNG1 can function not only as peptide:N-glycanase but also as transglutaminase
-
-
-
additional information
?
-
-
the Drosophila ortholog of PNGase (PngI) does not possess deglycosylation activity but retains carbohydrate-binding activity. The transglutaminase domain of Pngl participates in some type of catalytic activity even though the protein does not have the conventional deglycosylation activity
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATPase p97 + H2O
?
show the reaction diagram
Q9JI78
a cofactor-binding motif of p97 contained within the last 10 amino acid residues of the C terminus is both necessary and sufficient to mediate interactions of p97 with PNGase. Phosphorylation of p97s highly conserved penultimate tyrosine residue, which is the main phosphorylation site during T cell receptor stimulation, completely blocks binding of either PNGase or Ufd3 to p97. This observation suggests that phosphorylation of this residue modulates endoplasmic reticulum-associated protein degradation activity by discharging substrate-processing cofactors
-
-
?
class I MHC HC + H2O
?
show the reaction diagram
-
the enzyme is involved proteasomal degradation of glycosylated type I membrane protein class I MHC heavy chain, dislocated from ER to cytosol by cytomegalovirus-encoded glycoprotein US2 protein, overview
-
-
?
concanavalin A-precursor + H2O
?
show the reaction diagram
-
deglycosylation leads to conversion into an active lectin
-
-
?
N-glycoprotein + H2O
?
show the reaction diagram
-
PNGase is involved in the release of N-glycans from N-glycoproteins
-
-
?
Oryzias latipes glycophosphoprotein MU-1 + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11, yolk-absorptive stage
-
-
?
Oryzias latipes glycophosphoprotein MU-2 + H2O
?
show the reaction diagram
-
acid PNGase M from blastoderm stage 11, yolk-absorptive stage
-
-
?
proton-coupled folate transporter + H2O
?
show the reaction diagram
-
-
-
-
?
tyrosinase + H2O
?
show the reaction diagram
-
the enzyme is required for processing of a class I-restricted epitope from tyrosinase together with the cooperative action of endoplasmic reticulum aminopeptidase 1 and cytosolic protease, overview
-
-
?
melanopsin + H2O
?
show the reaction diagram
Q5XI55
-
-
-
?
additional information
?
-
-
involved in proteasomal degradation of misfolded glycoproteins
-
?
additional information
?
-
-
the cytosolic enzyme deglycosylates misfolded glycoproteins prior to the endoplasmic reticulum-associated protein degradation
-
-
-
additional information
?
-
-
the enzyme deglycosylates misfolded glycoproteins, the enzyme is a mediator for p97 functions, the PUB domain functions as a p97 binding module in human enzyme, p97 is an AAA ATPase with an ubiquitin-selective molecular machine involved in multiple cellular processes including protein degradation through the ubiquitin-proteasome system
-
-
-
additional information
?
-
-
the enzyme is responsible for deglycosylation of N-linked glycoproteins dislocated from endoplasmic reticulum to cytosol
-
-
-
additional information
?
-
-
the enzyme mediates the binding of the cytoplasmic proteins p97 and HR23B to the proteasome through formation of a ternary complex, p97 binds the autocrine motility factor receptor AMFR, modeling of interaction of the endoplasmic reticulum with the proteasome, overview
-
-
-
additional information
?
-
-
the enzyme removes N-linked oligosaccharides from misfolded glycoproteins as part of endoplasmic reticulum-associated degradation pathway involving a complex formation with proteins HR23B, cytosolic protein Y33K, p97, and autocrine motility factor receptor AMFR, the AAA ATPase p97 links peptide N-glycanase to the endoplasmic reticulum-associated E3 ligase AMFR, the N-terminus of PNGase interacts with the C-terminal tail of AMFR, complex formation model, overview
-
-
-
additional information
?
-
-
the enzyme removes N-linked oligosaccharides from misfolded glycoproteins as part of endoplasmic reticulum-associated degradation pathway involving a tight complex-formation with protein HR23, HR23 is also involved in DNA repair, co-evolution of the endoplasmic reticulum-associated degradation and DNA repair pathways, overview
-
-
-
additional information
?
-
-
SpPNGase deglycosylates the misfolded glycoproteins
-
-
-
additional information
?
-
-
the AtPNG1 gene encodes a bona fide peptide:N-glycanase that contributes to ERAD related protein quality control in plants
-
-
-
additional information
?
-
Q02890
catalyses the de-glycosylation of unfolded glycoproteins
-
-
-
additional information
?
-
D0QU16, -
the enzyme hydrolyses the beta-aspartyl-glycosylamine bond of N-linked glycoproteins/glycopeptides and releases free N-glycans
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
activity stimulating
Zn2+
-
zinc binding domain structure involving a CXXC motif, zinc binding stabilizes the enzyme conformation by stabilizing the intermediate state and promoting product release
Zn2+
-
zinc metalloenzyme
Zn2+
-
zinc metalloenzyme
Mn2+
-
partial activity stimulation
additional information
D0QU16, -
the enzyme does not require any metal ions for full enzymatic activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
benzyloxycarbonyl-VAD-fluoromethyl ketone
-
-
benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone
-
covalently bind to the active site Cys191 of recombinant yeast peptide N-glycanase
Ca2+
-
2 mM, 19% inhibition
carbobenzyloxy-Val-Ala-Asp
-
i.e. Z-VAD, binds to the active site of the enzyme, binding structure, overview
carbobenzyloxy-Val-Ala-Asp
-
i.e. Z-VAD, in vivo inhibition
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
i.e. Z-VAD-fmk, a broad-spectrum caspase inhibitor, binds covalently to the active site of the mammalian enzyme, in vivo inhibition of recombinant PNGase in U373 cells, overview
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
i.e. Z-VAD-fmk, a broad-spectrum caspase inhibitor, binds covalently to the active site of the mammalian enzyme
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
i.e. Z-VAD-fmk, a broad-spectrum caspase inhibitor, IC50: 0.05 mM, binds covalently to the active site of the yeast enzyme, no restoration of enzyme activity by dialysis
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
i.e. Z-VAD-fmk, in vivo inhibition
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
-
Co2+
-
28% remaining activity by 2 mM
Co2+
-
2 mM, almost complete inhibition
Cu2+
Prunus amygdalus var. dulcis
-
-
Cu2+
-
partial inhibition
Cu2+
-
2 mM, almost complete inhibition
CuSO4
-
nearly complete inhibition of the acid PNGase M by 5 mM, 10-20% remaining activity at neutral PNGase M by 5 mM
EDTA
-
23% inhibition by 5 mM, 13% inhibition by 10 mM
EDTA
-
activity is not restored by adding metal ions
EDTA
-
not restored by readdition of metal ions
Fbs1
-
Fbs1 captures malfolded glycoproteins, protecting them from the attack of PNGase, during the chaperoning or ubiquitinating operation in the cytosol
-
Fe2+
-
2 mM, 90% inhibition
-
Fe3+
Prunus amygdalus var. dulcis
-
-
-
FeCl3
-
nearly complete inhibition of the acid PNGase M by 5 mM, 10-20% remaining activity at neutral PNGase M by 5 mM
fucose residue alpha-1-3-linked or alpha-1-6-linked to the proximal GlnNAc residue
-
complete inhibition
-
Glycoasparagine
-
1 mM causes 86% inhibition
Glycoasparagine
-
-
K+
-
significant inactivation by 100 mM
L-hyosphorin derived nonapeptide
-
1 mM causes 92% inbition of acid PNGase M
-
liberated glycan reaction product
-
causes 82% inhibition
-
liberated glycan reaction product
-
-
-
Man8GlcNAc2-iodoacetoamide
-
-
Man9GlcNAc2-iodoacetoamide
-
strong inhibitor, irreversibly inhibits that catalytic Cys in a highly specific manner
Mg2+
-
2 mM, 14% inhibition
Mn2+
-
33% remaining activity by 2 mM
Mn2+
-
2 mM, 56% inhibition
Monoiodoacetic acid
-
2 mM causes 64% inhibition
Monoiodoacetic acid
-
5 mM causes 82% inhibition of the neutral PNGase M
N,N'-diacetylchitobiose
-
-
N,N'-diacetylchitobiose
-
GlcNAc2, competitive inhibitor may be directly accessible to the catalytic site
N,N'-diacetylchitobiose
Prunus amygdalus var. dulcis
-
-
N,N'-diacetylchitobiosylbromoacetamide
-
-
N,N'-diacetylchitobiosylchloroacetamide
-
-
N,N'-diacetylchitobiosyliodoacetamide
-
-
N-benzyloxycarbonyl-VAD-fluoromethylketone
Q02890
Z-VAD-fmk
N-ethylmaleimide
-
2 mM, significant inhibition by thiol-modification, alkylation
N-ethylmaleimide
-
5 mM causes complete inhibition
Na+
-
significant inactivation by 100 mM
p-chloromercuribenzoic acid
-
0.5 mM causes 89% inhibition of neutral PNGase M
phosphate buffers
-
no significant activity
-
SDS
-
complete inactivation by 0.05% SDS can be corrected by the addition of 1% Nonidet P-40
triomannose
-
-
triomannose
-
evidence for occupation of the carbohydrate-binding site
Tris-HCl
-
no significant activity
yeast mannan
-
-
-
Z-VAD-fmk
-
i.e. carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone, a broad-spectrum caspase inhibitor, potent inhibition, binding structure determination and analysis with wild-type and mutant C191A enzyme, the inhibitor binds covalently to the catalytic residue Cys191, but not to the catalytic His218
Zn2+
Prunus amygdalus var. dulcis
-
-
Zn2+
-
51% remaining activity by 2 mM
Zn2+
-
2 mM, almost complete inhibition
ZnCl2
-
10-20% remaining activity at neutral PNGase M by 5 mM
Monoiodoacetic acid
-
2 mM, significant inhibition by thiol-modification
additional information
-
no effect: yeast mannan, trimannose
-
additional information
Prunus amygdalus var. dulcis
-
no effect: thiol inhibitors, iodoacetamide, N-ethylmaleimide, actinomycete protease inhibitors, leupeptins, chymostatin, pepstatin, Mg2+, Ca2+, Mn2+, EDTA, L-cysteine, phenylmethylsulfonyl fluoride, gamma-D-gluconolactone, yeast mannan, triomannose
-
additional information
-
no effect: free peptides formed during reaction, mannose, alpha-methyl-D-mannoside, 5 mM Na2SO4
-
additional information
-
no effect: 1 mM peptide free N-linked glycan, 1 mM de-N-glycosylated peptide, 5 mM phenylmethylsulfonyl fluoride at neutral PNGase M; no effect: 5 mM monoiodoacetic acid, 0.5 mM p-chloromercurobenzoic acid, 5 mM ZnCl2 at acid PNGase M
-
additional information
-
the enzyme is preferably inhibited by aspartate-based inhibitors
-
additional information
-
no inhibition by the caspase inhibitor Q-VD-OPh
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
DTT
-
10 mM, absence causes inactivation of L-929 PNGase, suggesting S-H group essential for enzyme activity
DTT
-
mouse organ-derived PNGase requires at least 1 mM DTT for maximal activity
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.2
-
asialo fetuin glycopeptide I
-
-
-
0.871
-
asiolo-ovomucoid glycopeptide
-
-
-
1
-
Beta-aspartylglycosylamine
-
pH 5.5
4
-
bromelain glycopeptide
Prunus amygdalus var. dulcis
-
stem bromelain undecapeptide, glycopeptidase group C
-
2
-
bromelain undecapeptide
Prunus amygdalus var. dulcis
-
glycopeptidase group A
-
2.3
-
bromelain undecapeptide
Prunus amygdalus var. dulcis
-
glycopeptidase group B
-
0.114
-
Fetuin glycopeptide
-
-
-
0.525
-
Fetuin glycopeptide
-
-
-
1.46
-
Fetuin glycopeptide
Prunus amygdalus var. dulcis
-
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0046
-
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
-
0.05
-
carbobenzyloxy-Val-Ala-Asp-alpha-fluoromethylketone
-
i.e. Z-VAD-fmk, a broad-spectrum caspase inhibitor, IC50: 0.05 mM, binds covalently to the active site of the yeast enzyme, no restoration of enzyme activity by dialysis
0.0016
-
Man8GlcNAc2-iodoacetoamide
-
-
0.17
-
Man9GlcNAc2
-
-
0.0017
-
Man9GlcNAc2-iodoacetoamide
-
-
0.0028
-
N,N'-diacetylchitobiosylbromoacetamide
-
-
0.019
-
N,N'-diacetylchitobiosylchloroacetamide
-
-
0.0008
-
N,N'-diacetylchitobiosyliodoacetamide
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.00275
-
-
-
0.019
-
Prunus amygdalus var. dulcis
-
glycopeptidase group A
0.0538
-
-
di-dansyl fetuin glycopeptide
0.0623
-
Prunus amygdalus var. dulcis
-
glycopeptidase group C
0.0855
-
Prunus amygdalus var. dulcis
-
glycopeptidase group B
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
D0QU16, -
-
5
-
Prunus amygdalus var. dulcis
-
-
5
-
Prunus amygdalus var. dulcis
-
glycopeptidase group B/C
5.2
-
Prunus amygdalus var. dulcis
-
-
5.3
-
Prunus amygdalus var. dulcis
-
taka-amylase A
5.5
-
-
-
5.5
-
-
surface-displayed PNGase F
6
-
Prunus amygdalus var. dulcis
-
glycopeptidase group A
6.5
-
-
-
6.6
-
-
liver PNGase
6.6
-
-
using MES buffer
6.8
-
-
spleen PNGase
7
-
-
L-929 PNGase, brain PNGase, kidney PNGase
7
-
-
using HEPES buffer
7
-
-
the optimum deglycosylation pH of the Png1p-Rad23p complex is pH 7.0
7.2
-
-
assay at
7.2
-
-
assay at
7.2
-
Q02890
activity assay
7.5
-
-
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
7.5
Prunus amygdalus var. dulcis
-
-
3
5.5
-
-
3
6
D0QU16, -
enzyme activity declines rapidly above pH 6.0
3
8.6
-
80% of the maximum activity at pH 3.3, 65% of the maximum activity at pH 8.6
3.5
4
-
acid PNGase M, whole embryo, stage 18-28
4
7.5
-
pH 4.0: about 50% of maximal activity, pH 7.5: about 40% of maximal activity, surface-displayed PNGase F
4
8.5
-
-
5
10
-
pH 5: about 50% of maximal activity, pH 10.0: about 70% of maximal activity
6
9
-
pH 6: about 70% of maximal activity, pH 9.0: about 60% of maximal activity
6.5
7
-
no activity below 5.5
6.8
7
-
-
7.5
8.5
-
neutral PNGase M, middle blastula blastoderm, stage 7-13
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
-
-
-
22
-
-
room temperature
30
-
-
-
30
-
-
the optimum deglycosylation temperature of the Png1p-Rad23p complex is 30C
37
-
-
surface-displayed PNGase F
37
-
Q02890
activity assay
40
-
D0QU16, -
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
15
25
-
remaining activity 50% at 5C and 60% at 37C
20
37
-
20C: about 60% of maximal activity, 37C: about 55% of maximal activity
20
40
-
20C: about 80% of maximal activity, 40C: about 75% of maximal activity
20
45
-
20C: about 50% of maximal activity, 45C: about 60% of maximal activity, surface-displayed PNGase F
20
60
D0QU16, -
enzyme activity declines rapidly above 40C (80% activity at 50C, 55% activity at 60C, 30% activity at 70C, no activity at 80C)
25
37
-
-
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
the highest activities are found in the youngest parts of the plant: apical buds, flowers and leaf blades
Manually annotated by BRENDA team
-
recombinantly expressed in Escherichia coli
Manually annotated by BRENDA team
-
C3H mouse-derived L-929 fibroblast cells
Manually annotated by BRENDA team
-
the highest activities are found in the youngest parts of the plant: apical buds, flowers and leaf blades
Manually annotated by BRENDA team
Prunus amygdalus var. dulcis
-
-
Manually annotated by BRENDA team
-
decreases during fruit growth and ripening
Manually annotated by BRENDA team
-
the highest activities are found in the youngest parts of the plant: apical buds, flowers and leaf blades
Manually annotated by BRENDA team
additional information
-
the AtPNG1 gene is uniformly and constitutively expressed at low levels throughout all developmental stages of the plant
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35500
-
-
gel filtration
43000
-
-
gel filtration
66800
-
Prunus amygdalus var. dulcis
-
gel filtration
69000
-
-
gel filtration, native enzyme
73000
-
Prunus amygdalus var. dulcis
-
MALDI-MS, native enzyme
79500
-
-
estimation by HPLC, from a standard curve of proteins of known molecular mass
80000
-
-
gel filtration
150000
-
-
estimation for acid PNGase M by gel filtration
212000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 43208, MALDI mass spectrometry
?
-
x * 110000, SDS-PAGE
?
D0QU16, -
x * 65000, SDS-PAGE; x * 65800, calculated from amino acid sequence
?
-
x * 74000, SDS-PAGE
dimer
-
2 * 105000, homodimer, SDS-PAGE, 41% content of hydrophobic acids
dimer
-
-
dimer
Prunus amygdalus var. dulcis
-
-
dimer
-
537 amino acids, 59335 Da-translation product, about 70000 Da after processing, dissociation in SDS leads to heterodimer, cleaving between Thr 335 and Thr 336
dimer
Prunus amygdalus var. dulcis
-
1 * 55000 + 1 * 27000, SDS-PAGE and two-dimensional electrophoresis
monomer
-
1 * 35500, SDS-PAGE
monomer
-
1 * 34700, native enzyme, 314 amino acid open reading frame
monomer
Elizabethkingia meningoseptica, Prunus amygdalus var. dulcis
-
-
additional information
-
enzyme domain structure and enzyme-HR23 complex structure analysis, the enzyme contains a catalytic transglutaminase-like fold, a catalytic cleft structure, a zinc binding domain, and an XPCB association motif, overview
additional information
-
the PUB domain, comprising residues 11-109, functions as a p97 binding module in human enzyme, three-dimensional structure of the PUB domain
additional information
-
enzyme-Rad23 interaction and complex formation analysis, the UBA domain of Rad23 is required, rad mutant does not interact with the enzyme, overview
additional information
-
the N-terminus of PNGase interacts with the C-terminal tail of AMFR, and PNGase interacts with protein p97 essentially requiring the enzymes' PUB domain, overview, complex formation model, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
59335 primary translation product + 9 to 11 posttranscriptional added high mannose chains, average Man5GlcNAc2
glycoprotein
D0QU16, -
the enzyme possesses 10 N-glycosylation sites at positions of 220, 266, 386, 397, 447, 481, 489, 551, 556 and 570 in the deduced polypeptide chain
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified wild-type PUB domain, also as selenomethionine variants, by sitting drop vapour diffusion method at 17C, with a reservoir solution containing 0.2 M sodium acetate, 0.1 M Tris, pH 8.5, 30% PEG 4000, using 20% glycerol as cryoprotectant, X-ray diffraction structure determination and analysis at 1.6 A resolution, mutant L66M/L75M/L87M PUB domain are crystallized in 50% PEG 400, 0.1 M CHES, pH 9.5, 0.2 M NaCl, molecular replacement, X-ray diffraction structure determination and analysis at 1.9 A resolution
-
hanging drop vapor diffusion, crystal structure of the N-terminal domain of PNGase in complex with the cofactor-binding motif of p97 contained within the last 10 amino acid residues of the C terminus provides detailed insight into the interaction between p97 and its substrate-processing cofactors
-
purified recombinant enzyme core domain and XPCB domain in complex with the recombinant murine HR23B protein, 9.5 mg/ml of enzyme domains in a 1:1 ratio, vapour diffusion method, against reservoir solution containing 0.1 M Tris-HCl, pH 8.5, 28-32% PEG 4000, 0.2 M sodium acetate, heavy atom derivatization by soaking in 1 mM ethyl mercury thiosalicylate, 1 mM K2[PtCN4], or 1 mM KAuCN2 for 4 h, cryoprotection by 20% glycerol, X-ray diffraction structure determination and analysis at 1.85 A resolution
-
the crystal structure of PNGase in complex with N,N'-diacetylchitobiose is described, refined at 3.4 A
Q02890
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
stable up to 24 h, unstable at temperatures above 30C after 1-3 h
37
45
-
Png1p is inactive at 37C. In contrast, the Png1p-Rad23p complex still possesses enzymatic activity at 45C
37
-
-
long periods in absence of detergent
37
-
-
rapidly reduced stability above 37C
62
-
-
incubation for 8 h retains 65% of the original activity
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
zinc binding stabilizes the enzyme conformation by stabilizing the intermediate state and promoting product release
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
readily inactivated by SDS
-
288925
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
active fractions stored at -70C
-
2 months at -20C, activity loss less than 50%
Prunus amygdalus var. dulcis
-
several weeks at 4C
Prunus amygdalus var. dulcis
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
native enzyme by Fe2+ affinity and phosphopeptide affinity chromatography to over 95% purity
-
recombinant wild-type and mutant enzyme fusion PUB domain from Escherichia coli strain C41(DE3) by nickel affinity chromatography, ion exchange chromatography, and gel filtration
-
460fold
-
glutathione-Sepharose column chromatography
-
recombinant full-length enzyme, enzyme core domain, and enzyme XPCB domain from Escherichia coli strain BL21(DE3),the XPCB domain by chitin affinity chromatographyand gel filtration
-
recombinant GST-tagged proteasome components HR23B, S4, and AMFR from Escherichia coli by glutathione affinity batch method, recombinant His6-tagged wild-type enzyme and truncated variants from Escherichia coli by nickel affinity chromatography
-
-
Prunus amygdalus var. dulcis
-
14500fold
Prunus amygdalus var. dulcis
-
recombinant wild-type and mutant Png1 from Escherichia coli strain BL21(DE3)
-
Ni-nitrilotriacetate column chromatography
D0QU16, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Saccharomyces cerevisiae
-
importantly, heterologous expression of AtPNG1 restores N-glycanase activity in a PNGase-deficient Saccharomyces cerevisiae mutant
-
expression in Escherichia coli and CHO cells
-
expressed in Sf21 insect cells
-
expression in Escherichia coli
-
the PNGase F gene from Flavobacterium meningosepticum is cloned into plasmid pYD1, which enables regulated expression, secretion and detection. The expression of PNGase F gene at extracellular surface of Saccharomyces cerevisiae is confirmed by immunofluorescence microscopy. Fluorescence activated cell sorter analysis indicates that, after 36 h cultivation, 47.6% of the cell surface is anchored with target proteins. The surface engineered enzyme is confirmed to be active and reached its highest level after induced for 36 h
-
expression of the enzyme in U373 astrocytoma cells
-
expression of wild-type and mutant enzyme PUB domain, comprising residues 11-109, fused to the lipoyl domain of Bacillus stearothermophilus dehihydrolipoamide acetyltransferase in Escherichia coli strain C41(DE3)
-
co-expression of His-tagged or GST-tagged truncated enzyme variants and of GST-tagged enzyme mutants with His-tagged p97, transient co-expression and complex formatin of N-terminal GFP-tagged PNGase and c-myc-tagged AMFR in COS-1 cells
-
co-expression of the His6-tagged wild-type enzyme and truncated variants with GST-tagged murine proteasome components HR23B, S4, and autocrine motility factor receptor AMFR in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) Codon-Plus cells
-
expression of HA-tagged wild-type and mutant enzymes in U373 astrocytoma cells
-
overexpression of the full-length enzyme, enzyme core domain, and enzyme XPCB domain in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli BL21 cells
-
co-expression of His6-tagged full-length enzyme or truncated versions with full-length Rad23 Escherichia coli strain BL21(DE3)
-
expression in Escherichia coli as inclusion bodies
-
expression of His- or FLAG-tagged wild-type and mutant enzymes, co-expression with Rad23
-
gene png1, expression of wild-type and mutant Png1 in Escherichia coli strain BL21(DE3)
-
into the vector pET28a for expression in Escherichia coli BL21DE3 Codon Plus RIL cells
Q02890
expression in Escherichia coli
-
expressed in Escherichia coli strain JM109 and in Pichia pastoris strain GS115
D0QU16, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C251A
-
the mutant exhibits a defect in degrading RTADELTA-transmembrane-Leu2 protein. Expression of the catalytic mutant results in significant stabilization of RTADELTA protein by binding to N-glycans on the substrate
C306A
-
site-directed mutagenesis, inactive mutant, inhibitor Z-VAD-fmk does not bind to the mutant enzyme
N41P
-
site-directed mutagenesis, the mutation completely abolishes the interaction of PNGase with p97
N58A
-
site-directed mutagenesis, the mutation does not affect the interaction of PNGase with p97
A208C
-
inactive
A208C/Y235H
-
inactive
Y235H
-
inactive
C165T
-
inactive
C165T/N166V/R167C
-
inactive
C191A
-
no enzymic activity
C191A
-
site-directed mutagenesis, inactive mutant, inhibitor Z-VAD-fmk does not bind to the mutant enzyme
C191A
-
site-directed mutagenesis, inactive mutant
C191A
-
mutant enzyme does not bind to the inhibitor Man9GlcNAc2-iodoacetoamide
C191A
-
catalytically inactive mutant enzyme
C191A
-
inactive
C221T
-
mutant with more than 50% wild type activity
D179E/P180A
-
mutant with more than 50% wild type activity
D208R/V209A
-
mutant with 10-50% wild type activity
D217A
Q02890
mutation in the peptide binding site
D235A
-
no enzymic activity
D235A
-
site-directed mutagenesis, inactive mutant
E185R
-
mutant with 10-50% wild type activity
E185R/T186V
-
mutant with 10-50% wild type activity
E193D
-
inactive
E193D/W194F
-
inactive
E222A
-
no enzymic activity
E238A
Q02890
mutation in the chitobiose binding site
F224Y
-
mutant with more than 50% wild type activity
G206D/L207I
-
mutant with 10-50% wild type activity
H218A
-
no enzymic activity
H218A
-
site-directed mutagenesis, inactive mutant
H218A
Q02890
mutation in the chitobiose binding site
H218F
Q02890
mutation in the chitobiose binding site
H218Y
-
site-directed mutagenesis, the mutant is inactive in protein glycosylation, but interacts with protein Rad23
I181W
-
mutant with more than 50% wild type activity
I181W/K182Q
-
inactive
K182Q
-
mutant with more than 50% wild type activity
K253A
Q02890
mutation in the chitobiose binding site
L198V
-
mutant with more than 50% wild type activity
L200M
-
mutant with more than 50% wild type activity
N166V/R167C
-
the mutant shows 79% of wild type activity
N178A
Q02890
mutation in the peptide binding site
N178T
-
mutant with 10-50% wild type activity
N214C/R215Q
-
mutant with 10-50% wild type activity
N266F/F227H
-
mutant with more than 50% wild type activity
Q239A
Q02890
mutation near the nonreducing end of the chitobiose, possible mannose binding site
Q243A
Q02890
mutation near the nonreducing end of the chitobiose, possible mannose binding site
R176A
Q02890
mutation in the peptide binding site
R187K/K188R
-
mutant with more than 50% wild type activity
R210A
-
no enzymic activity
V219L
-
mutant with 1-10% wild type activity
W123A
Q02890
mutation in the peptide binding site
W194F/C195A
-
mutant with 10-50% wild type activity
W220F
-
81% of activity compared to wild type
W231F
-
83% of activity compared to wild type
W251A
Q02890
mutation in the chitobiose binding site
Y211W
-
mutant with more than 50% wild type activity
L66M/L75M/L87M
-
site-directed mutagenesis, the PUB domain mutant shows slightly reduced p97 binding
additional information
-
construction of PNGase U-373MG siRNA cell lines, the glycosylated type I membrane protein class I MHC HC, dislocated by US2 protein, becomes cytosolic when peptide: N-glycanase is compromised, overview
G79/F80A
-
site-directed mutagenesis, the double mutation completely abolishes the interaction of PNGase with p97
additional information
-
construction of truncated enzyme variants comprising amino acid residues 1-171, 1-471, 471-651, 171-471, 1-111, 112-450, 451-651, and 1-450
additional information
-
construction of truncated enzyme variants comprising amino acid residues 1-80, 35-81, 1-181, 1-181DELTAPUB, 1-130, 1-111, 112-450, and 451-651
L200M/I201L
-
mutant with more than 50% wild type activity
additional information
-
enzyme deletion strain, formation of free oligosaccharides is reduced by 70-80%
additional information
-
site-directed mutagenesis of not conserved amino acids
additional information
-
deletion of the N-terminal H1 helix (Png1p-DH1) enhances the deglycosylation activity of peptide:N-glycanase towards denatured glycoproteins
Y223I
-
mutant with 10-50% wild type activity
additional information
-
deletion of the N-terminal H1 helix (Png1p-DH1) enhances the deglycosylation activity of peptide:N-glycanase towards denatured glycoproteins
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
valuable tool to characterize the peptide portion of glycoproteins and glycopeptides to release asparagine-linked oligosaccarides for structural analysis, recombinant PNGase F contains no contaminant Endo F, proteases or exoglycosidases
analysis
-
a procedure to map N-glycosylation sites is presented, it can be applied to purified proteins as well as to highly complex mixtures. The method exploits deglycosylation by PNGase F in a diagonal, reverse-phase chromatographic setup
synthesis
-
PNGase F can be utilized for glycosylation of non-glycosylated recombinant proteins produced in prokaryotic cells