Information on EC 3.4.22.65 - peptidase 1 (mite)

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The expected taxonomic range for this enzyme is: Dermatophagoides

EC NUMBER
COMMENTARY
3.4.22.65
-
RECOMMENDED NAME
GeneOntology No.
peptidase 1 (mite)
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
broad endopeptidase specificity
show the reaction diagram
-
-
-
-
the proteolytic activity of Der f 1 is a major contributor to its allergenicity. Cysteine protease activity of Der p 1 enhances total IgE production
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
allergen Der f 1
-
-
allergen Der f1
-
-
-
-
CO1.073
-
-
-
-
Der f 1
-
-
-
-
Der f 1
Q58A71
-
Der p 1
-
-
-
-
Der p 1
-
automaturation product of proDer p 1 (proDer p 1.0105 isoform)
dust mite allergen Der p 7
P49273
-
house dust mite allergen
-
-
house dust mite allergens
-
HDM
mite major group 1 allergens
-
-
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
the isozymes have allergenic potential in humans
physiological function
-
Der p 7 elicits strong IgE antibody and T-cell responses in mite allergic patients. Sensitization to house dust mite allergens is strongly correlated with asthma. The allergen function may contribute to allergenicity, e.g. the protease activity of Group 1 mite allergens, and the interaction with the innate immune system by Group 2 mite allergens
additional information
-
the Group 7 as well as the Group 2 mite allergens are structurally similar to different proteins in the TLR pathway
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2-aminobenzoyl)-Val-Ala-norLeu-Ser-Tyr(3-NO2)-Asp + H2O
?
show the reaction diagram
-
ADZ 50,059
-
-
?
2-aminobenzoic acid-Val-Ala-Nle-Ser-(3-nitro)-tyrosinyl-aspartamide + H2O
?
show the reaction diagram
-
-
-
-
?
alpha1-antitrypsin + H2O
?
show the reaction diagram
-
recombinant Der p 1 previously activated with L-cysteine or DTT
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
Azocoll + H2O
?
show the reaction diagram
-
-
-
-
?
Azocoll + H2O
?
show the reaction diagram
-
collagen substrate
-
-
?
Azocoll + H2O
?
show the reaction diagram
-
collagen substrate, Der f 1-N53Q proteolytically active
-
-
?
butyloxycarbonyl-Gln-Ala-Arg-4-methylcoumaryl-7-amide + H2O
butyloxycarbonyl-Gln-Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Gln-Ala-Arg-7-amido-4-methylcoumarin + H2O
butyloxycarbonyl-Gln-Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Gln-Ala-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Gln-Ala-Arg-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin
show the reaction diagram
-
short synthetic substrate
-
-
?
butyloxycarbonyl-Gln-Ala-Arg-MCA + H2O
butyloxycarbonyl-Gln-Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Gln-Gly-Arg-MCA + H2O
butyloxycarbonyl-Gln-Gly-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Phe-Ser-Arg-MCA + H2O
butyloxycarbonyl-Phe-Ser-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
more efficiently hydrolyzed than N-succinyl-Ala-Pro-Ala-7-amido-4-methylcoumarin
-
-
?
butyloxycarbonyl-Val-Leu-Lys-MCA + H2O
butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
CD23 + H2O
?
show the reaction diagram
-
-
-
-
?
CD23 + H2O
?
show the reaction diagram
-
CD23 is a calcium-dependent type II integral membrane protein. CD23 from the surface of cultured human B cells is cleaved at two sites: Ser155-Ser156 and Glu298-Ser299 to produce a 17000 Da fragment containing the lectin domain and only part of the C-terminal tail. No such effect is demonstrable with mouse CD23
-
-
?
CD23 + H2O
?
show the reaction diagram
-
cleaves CD23 from the surface of cultured human B cells. The cleavage of the receptor from the B cell sulface is associated with a parallel increase in soluble CD23, Der p 1 in addition to being highly immunogenic may up-regulate IgE synthesis by virtue of its ability to cleave CD23
-
-
?
CD23 + H2O
?
show the reaction diagram
-
low-affinity IgE receptor
-
-
?
CD25 + H2O
?
show the reaction diagram
-
-
-
-
?
CD25 + H2O
?
show the reaction diagram
-
alpha-subunit of the IL-2 receptor
-
-
?
chestnut cystatin + H2O
?
show the reaction diagram
-
specific proteolytic cleavage between Gly6 and Val7, giving rise to a noninhibitory processed protein. Not cleaved by Der f 1 from Dermatophygoides farinae
-
-
?
Der p 1 propiece + H2O
?
show the reaction diagram
-
potent competitive inhibitor of Der p 1. The Der p 1 propiece behaves as a substrate and is fully degraded during this interaction. The rapid inactivation of Der p 1 prodomain is a mechanism that may contribute to the potency of this allergen
-
-
?
human alpha1-antitrypsin + H2O
?
show the reaction diagram
-
cleaved by Der f 1-N53Q but not without its activation
-
-
?
human CD23 + H2O
?
show the reaction diagram
-
cleaved by Der f 1-N53Q and natural Der f 1 on the cell surface in a time- and dose dependent manner
-
-
?
human CD23 + H2O
?
show the reaction diagram
-
human low-affinity IgE receptor
-
-
?
human CD25 + H2O
?
show the reaction diagram
-
alpha subunit of the human IL-2 receptor
-
-
?
human CD25 + H2O
?
show the reaction diagram
-
cleaved by Der f 1-N53Q and natural Der f 1 on the cell surface in a time- and dose dependent manner
-
-
?
lung surfactant protein A + H2O
?
show the reaction diagram
-
-, degradation of lung surfactant protein A and lung surfactant protein D is associated with diminished binding to carbohydrates and to Dermatophagoides pteronyssinus allergen 1 itself and diminishes capacity to agglutinate bacteria. The degradation and consequent inactivation of lung surfactant protein A and lung surfactant protein D may be a novel mechanism to account for the potent allergenicity of the dust mite allergen
-
-
?
N-Bz-Phe-Val-Arg-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-succinyl-Ala-Pro-Ala-7-amido-4-methylcoumarin
N-succinyl-Ala-Pro-Ala + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
N-succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin
N-succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
more efficiently hydrolyzed than N-succinyl-Ala-Pro-Ala-7-amido-4-methylcoumarin
-
-
?
N-tert-butoxycarbonyl-Gln-Ala-Arg-7-amido-4-methylcoumarin + H2O
N-tert-butoxycarbonyl-Gln-Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
succinyl-Leu-Leu-Val-Tyr-MCA + H2O
succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Z-Phe-Arg-4-methylcoumarin 7-amide + H2O
Z-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
P49273
-
-
-
?
lung surfactant protein D + H2O
?
show the reaction diagram
-
-, degradation of lung surfactant protein A and lung surfactant protein D is associated with diminished binding to carbohydrates and to Dermatophagoides pteronyssinus allergen 1 itself and diminishes capacity to agglutinate bacteria. The degradation and consequent inactivation of lung surfactant protein A and lung surfactant protein D may be a novel mechanism to account for the potent allergenicity of the dust mite allergen
-
-
?
additional information
?
-
-
mature recombinant enzyme shows the same IgE binding activity as the native enzyme
-
-
-
additional information
?
-
-
allergen from Dermatophagoides farinae
-
-
-
additional information
?
-
-
Der p 1 is the most immunodominant allergen involved in the expression of dust mite-specific immunoglobulin (Ig)E-mediated hypersensitivity. The proteolytic activity of Der p 1 is a major contributor to its allergenicity. Cysteine protease activity of Der p 1 enhances total IgE production
-
-
-
additional information
?
-
-
cysteine protease activity of Der p 1 enhances total IgE production, apart from increasing Der p 1-specific IgE. This allergen may play a central role in destabilizing the micro-environment within target tissues to one that is pro-allergic, and thus aid in the initiation and propagation of the allergic cascade
-
-
-
additional information
?
-
-
recombinant Der f 1 activated with L-cysteine reduces the barrier function of the skin in dose- and time-dependent manners. The reduction is dependent on its proteolytic activity
-
-
-
additional information
?
-
-
the efficient in vivo responses, including production of IgE and IgG against the highly purified rDer p 1, are dependent on the cysteine protease activity in mice. The three types of rDer p 1 differing in function or structure elicit distinctly different immune responses
-
-
-
additional information
?
-
-
the house dust mite allergen Der p 1 stimulates the expression of interleukin-8 in human airway epithelial cells via a proteinase-activated receptor-2-independent mechanism
-
-
-
additional information
?
-
-
house dust mite major allergens Der p 1 and Der p 5 activate human airway-derived epithelial cells by protease-dependent and protease-independent mechanisms
-
-
-
additional information
?
-
-
Der p 7 binds a bacterially-derived lipid product, a common feature of some allergens, ligand binding analysis by NMR analysis
-
-
-
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
CD23 + H2O
?
show the reaction diagram
-
Der p 1 in addition to being highly immunogenic may up-regulate IgE synthesis by virtue of its ability to cleave CD23
-
-
?
lung surfactant protein A + H2O
?
show the reaction diagram
-
degradation of lung surfactant protein A and lung surfactant protein D is associated with diminished binding to carbohydrates and to Dermatophagoides pteronyssinus allergen 1 itself and diminishes capacity to agglutinate bacteria. The degradation and consequent inactivation of lung surfactant protein A and lung surfactant protein D may be a novel mechanism to account for the potent allergenicity of the dust mite allergen
-
-
?
lung surfactant protein D + H2O
?
show the reaction diagram
-
degradation of lung surfactant protein A and lung surfactant protein D is associated with diminished binding to carbohydrates and to Dermatophagoides pteronyssinus allergen 1 itself and diminishes capacity to agglutinate bacteria. The degradation and consequent inactivation of lung surfactant protein A and lung surfactant protein D may be a novel mechanism to account for the potent allergenicity of the dust mite allergen
-
-
?
additional information
?
-
-
allergen from Dermatophagoides farinae
-
-
-
additional information
?
-
-
Der p 1 is the most immunodominant allergen involved in the expression of dust mite-specific immunoglobulin (Ig)E-mediated hypersensitivity. The proteolytic activity of Der p 1 is a major contributor to its allergenicity. Cysteine protease activity of Der p 1 enhances total IgE production
-
-
-
additional information
?
-
-
cysteine protease activity of Der p 1 enhances total IgE production, apart from increasing Der p 1-specific IgE. This allergen may play a central role in destabilizing the micro-environment within target tissues to one that is pro-allergic, and thus aid in the initiation and propagation of the allergic cascade
-
-
-
additional information
?
-
-
recombinant Der f 1 activated with L-cysteine reduces the barrier function of the skin in dose- and time-dependent manners. The reduction is dependent on its proteolytic activity
-
-
-
additional information
?
-
-
the efficient in vivo responses, including production of IgE and IgG against the highly purified rDer p 1, are dependent on the cysteine protease activity in mice. The three types of rDer p 1 differing in function or structure elicit distinctly different immune responses
-
-
-
additional information
?
-
-
the house dust mite allergen Der p 1 stimulates the expression of interleukin-8 in human airway epithelial cells via a proteinase-activated receptor-2-independent mechanism
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
dithiothreitol dependent proteolytic activity
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
a flexible metal binding site is present in pro-Der p 1
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-(2-aminoethyl) benzenesulfonyl fluoride
-
-
chestnut cystatin
-
inhibits the enzyme and greatly increases the larval mortality
-
cystatin A
-
sweat inhibitor, upregulation of IL-8 secretion by stimulation of human keratinocytes by Der f 1 is blocked
-
cystatin A
-
-
-
Der p 1 allergen
-
ADZ 50,000, irreversible inactivation, active site titrant
-
Der p 1 prodomain
-
PP-02, recombinant, apparently competitive
-
Der p 1 propiece
-
potent competitive inhibitor of Der p 1. The Der p 1 propiece behaves as a substrate and is fully degraded during this interaction
-
E-64
-
irreversible inhibition, cysteine protease-specific
E-64
-
trans-epoxysuccinyl L-leucylamido (4-guanidine) butane, 0.01 mM, complete inhibition of interleukine production in A549 epithelial cells
E-64
-
cysteine protease specific inhibitor
E-64
-
cysteine protease-specific inhibitor, complete inhibition
E-64
-
cysteine protease-specific inhibitor
E-64
-
cysteine protease-specific inhibitor, 0.1 mM, complete inhibition
egg-white cystatin
-
-
-
phenylmethanesulfonyl fluoride
-
-
PTL11028
-
a highly potent and specific inhibitor, being effective against both purified protease and Der p 1 within HDM extract
-
additional information
-
Der f1 from Dermatophagoides farinae is highly susceptible, Der p 1 from Dermatophagoides pteronyssinus is not affected by chestnut cystatin. Der p 1 inactivates chestnut chystatin by a specific proteolytic cleavage between Gly6 and Val7
-
additional information
-
not inhibited by AEBSF
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
cysteine
-
the enzyme is completely dependent on preactivation with cysteine
additional information
-
Der p 1 dose-dependently increase the production of IL-6 and IL-8, can be blocked by heat treatment and cell desquamation
-
additional information
-
Der p 1 induce the release of IL-1beta, IL-6, IL-10, tumor necrosis factor-alpha and granulocyte macrophage colony-stimulating factor from eosinophils, surface expression of CD18 and ICAM-1, can activate NK-KB, AP-1 activity and p38 MAPK in BEAS-2B cells and eosinophils alone and in co-culture
-
additional information
-
Der p 1 stimulates the expression of Interleukin-8 in human pulmonary epithelial cells via the ERK1/2 signaling pathway
-
additional information
-
Der p 1 induces chemokine production (CXCL8, CXCL10, CCL2, CCL5 and CCL20) by bronchial epithelial cells BEAS-2B and BECs from nonatopic controls through protease-activated receptor (PAR)-2 cleavage
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0154
-
(2-aminobenzoyl)-Val-Ala-norLeu-Ser-Tyr(3-NO2)-Asp
-
Der p 1 purified from Dermatophagoides pteronyssinus
14.4
-
(2-aminobenzoyl)-Val-Ala-norLeu-Ser-Tyr(3-NO2)-Asp
-
recombinant Der p 1, expressed in Pichia pastoris
0.003
-
2-aminobenzoic acid-Val-Ala-Nle-Ser-(3-nitro)-tyrosinyl-aspartamide
-
pH 8.3
0.017
-
casein
-
pH 6.5, 37C
0.033
-
casein
-
pH 6.5, 30C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.2
-
2-aminobenzoic acid-Val-Ala-Nle-Ser-(3-nitro)-tyrosinyl-aspartamide
-
pH 8.3
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000031
-
chestnut cystatin
-
pH 6.5, 30C
-
0.000000005
-
cystatin A
-
pH 8.3
-
0.0000279
-
cystatin C
-
pH 8.3
-
0.00126
-
cystatin D
-
pH 8.3
-
0.0000148
-
cystatin E/M
-
pH 8.3
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000051
-
Der p 1 prodomain
-
recombinant Der p 1, expressed in Pichia pastoris
-
0.0000569
-
Der p 1 prodomain
-
Der p 1 purified from Dermatophagoides pteronyssinus
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.66
-
Q58A71
calculated
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
wild-type pro-Der f1 is secreted to culture medium of baculovirus-transfeceted SF9 cells, but pro-N53Q is not
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
17000
18000
-
recombinant form without N-glycosylation, gel filtration
17000
-
-
Der f 1-N53Q, gel filtration
18000
-
-
natural Der f 1, gel filtration
25000
-
-
nature form, gel filtration
25000
-
-
gel filtration, Der p 1 purified from Dermatophagoides pteronyssinus
36000
60000
-
smear from 36 to 60 kDa, SDS-PAGE, recombinant Der p 1, expressed in Pichia pastoris
55000
-
-
average molecular weight, gel filtration, recombinant Der p 1, expressed in Pichia pastoris
additional information
-
-
recombinant Der p 1 and Der p 1 purified from Dermatophagoides pteronyssinus show differences in glycosylation
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 34000-45000, mature enzyme, SDS-PAGE
?
-
x * 24000,mature mutant enzyme N52Q, SDS-PAGE under non-reducing conditions; x * 35000-45000, mature wild-type enzyme, SDS-PAGE under non-reducing conditions; x * 35000-45000, mature wild-type enzyme under reducing conditions; x * 40000-50000, mature mutant enzyme N52Q, SDS-PAGE under non-reducing conditions
?
-
x * 28000, SDS-PAGE
?
Q58A71
x * 34000, SDS-PAGE
?
-
x * 29000, ProDer p1, SDS-PAGE; x * 35000, recombinant ProDer p1 (Escherichia coli), SDS-PAGE
?
-
x * 22200, about, mature Der p 7, sequence calculation, x * 24000-31000, SDS-PAGE
monomer
-
1 * 17000-18000, gel filtration and SDS-PAGE
monomer
-
SDS-PAGE
monomer
-
recombinant ProDer p 1 (CHO cell), gel filtration
monomer
-
gel filtration, pH 7.5
monomer
-
crystal structure: analysis of the accessible surface area upon dimerization, dimer corresponds most probably to the inactive form
additional information
-
Der p 7 has an elongated structure with two 4-stranded anti-parallel beta-sheets which wrap around a long C-terminal helix, NMR analysis of His-tagged Der p 7, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
glycoprotein
-
the wild-type recombinant enzyme is more highly glycosylated than the native enzyme
glycoprotein
-
GlcNAc is contained at the non-reducing terminus of the sugar chains of native Der f1 but not of lysylendopeptidase-treated reDEr f1 E(-1)K. Lysylendopeptidase-treated reDEr f1 E(-1)K reacts with Lens culinaris agglutinin but native Der f1 does not , suggesting that fucose is connected with GlcNAc, bound to an asparagine residue, but not of native Der f1
glycoprotein
-
N-glycosylation is essential for secretion in insect Sf9 cells but not in Pichia pastoris
proteolytic modification
-
the proforms of Der p 1 and Der f1 differ in the N-terminal sequences
proteolytic modification
-
the N-terminal sequences of the secreted proteins are determined to start from -59 or -78, downstream from the putative signal-cleavage site in the prosequence of Der f 1. Those of processed proteins are determined to start from -2 or 1, two residues upstream from or at the N-terminus of native Der f 1. The prosequence is removed autocatalytically by dialysis against acidic buffer
proteolytic modification
-
the presence of the pro-sequence is inhibitory to the IgE-binding activity of Der f1
proteolytic modification
-
the processed reDer f1 E(-1)K can be obtained after lysylendopeptidase and endoglycosidase Hf treatment
proteolytic modification
-
the removal of the pro-sequence is necessary for the cysteine protease activity. Acid treatment of the renatured pro Der f1 results in the autocatalytic removal of the pro-sequence. The obtained mature form of Der f1 binds IgE in patient sera and induces the release of histamine from peripheral blood leukocytes equally to native Der f1
glycoprotein
-
N-glycosylation of the prosequences decelerated the maturation. Systems using Pro-Der p 1 without the prodomain glycosylation are useful for the efficient preparation of a recombinant mature allergen
glycoprotein
-
different levels of glycosylation or isozymes
proteolytic modification
-
contribution of the N-terminal region of the Der p 1 prosequence including the N-glycosylation site, Asn(-65), on effective inhibition of proteolytic activity in pro-Der p 1
proteolytic modification
-
the Der p 1 propiece is a potent competitive inhibitor of Der p 1. The Der p 1 propiece behaves as a substrate and is fully degraded during this interaction. The rapid inactivation of Der p 1 prodomain is a mechanism that may contribute to the potency of this allergen
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, 45% monomethyl polyethylene glycole 5000 or 2000 in 50 mM sodium acetate (pH 5.5), 1.9 A resolution, typical papain-family fold, exhibits a magnesium binding site
-
recombinant Der p 7-MBP fusion protein, mixing of 0.001 ml of 27 mg/mL protein in 25 mM HEPES, pH 7.4, 75 mM NaCl, and 5 mM maltose, with 00.001 ml of reservoir solution containing 42.5 mM Tris pH 8.5, 12.75% PEG 4000, 85 mM lithium sulfate, and 7.5% glycerol, 4C, 1 week, X-ray diffraction structure determination and analysis at 2.35 A resolution, molecular replacement
-
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
most of the reDer f1 E(-1)K produced by the transformants is degraded when culture is shaken vigorously. The degradation of reDer f1 E(-1)K is suppressed when it is shaken gently
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
column chromatography
-
mature form purified by anion exchange and gel filtration chromatography
-
mature forms were purified by anion-exchange and gel filtration chromatography, proforms were purified by means of the same methods as the mature forms but without activation by dialysis against acidic buffer, natural Der f 1 was purified from whole culture extracts of Dermatophagoides farinae
-
affinity chromatography
-
ammonium sulfate precipitation, gel filtration, ion exchange chromatography
-
anion exchange and gel filtration
-
culture supernatants recovered, buffer exchange by gel filtration, equilibration with 100 mM sodium acetate buffer (pH 4)
-
dialysis, cationic exchange on SP-Sepharose, recombinant Der p 1 dialysis, benzamidine column, dialysis
-
immunoaffinity chromatography using immobilized 4C1 anti-Der p 1 monoclonal antibody
-
proform secreted in Pichia pastoris culture supernatant and converted into their prosequence-removed mature form, anion exchange column and gel filtration
-
recombinant Der p 7-MBP fusion protein Der p 7 from Escherichia coli Rosetta2(DE3) pLacI cells by amylose affinity chromatography and gel filtration. Recombinant His6-tagged Der p 7 from Escherichia coli Rosetta2(DE3) pLacI cells by alternating nickel affinity and anion exchange chromatography
-
recombinant His-tagged fusion proteins by nickel affinity chromatography using a His-Trap column under denaturing conditions with 20 m M sodium phosphate, pH 7.0, 0.5 M NaCl, 0.05% Tween 20, 8 M urea, 1 mM DTT, 0.03 M imidazole as binding buffer, and 20 mM sodium phosphate, pH 7.8, 0.05% Tween 20, 8 M urea, 1 mM DTT, 1 M imidazole as elution buffer
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
construction of an isopropyl-beta-D-thiogalactopyranoside-inducible expression plasmid to produce the pro-form of Der f1 in Escherichia coli
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DNA fragment encoding precursor-type recombinant Der f1 E(-1)K, which has the C-terminal amino acid of the pro-sequence, Glu, changed to Lys, expression in Aspergillus oryzae
-
expressed in Saccharomyces cerevisiae
-
production of recombinant enzyme in insect cells using a baculovirus expression system
-
recombinant Der f1 that is produced using a baculovirus expression sytem contains pro-sequences of different length. Most of theses can be removed by acid treatment. The N-terminal amino acids of the acid-treated recombinant enzyme are uniform. A method for processing of the pro-sequence has been developed by producing reDer f1 E(-1)K with baculovirus expression system in which the carboxy terminal amino acid of the pro-sequence (glutamate) is replaced by lysine. Addition of lysylendopeptidase into the culture medium leads to processing of the pro-sequence of reDer f1 E(-1)K
-
recombinant proforms were secreted into the culture of Pichia pastoris transfectant cells and converted into the prosequence-removed mature forms by means of dialysis against an acidic buffer
-
the preproforms of Der f 1 and a mutant N53Q, whose consensus motif for N-glycosylation is disrupted, are expressed in Pichia pastoris
-
wild-type pro-Der f1 is secreted to culture medium of baculovirus-transfeceted SF9 cells, pro-N53Q is not secreted in insect Sf9 cells although secreted in Pichia pastoris
-
Der p 1.0124 (according to the IUIS-WHO Allergen Nomenclature Subcommittee)
-
expressed in Pichia pastoris
-
expression and maturation of prepro-Der p 1 and its mutants in Pichia pastoris. Maturation systems using Pro-Der p 1 without the prodomain glycosylation are useful for the efficient preparation of a recombinant mature allergen; expression in Pichia pastoris
-
expression in Escherichia coli
-
expression in Pichia pastoris X-33
-
expression of Der p 7-MBP fusion protein and His6-tagged Der p 7 in Escherichia coli Rosetta2(DE3) pLacI cells
-
expression of His-tagged fusion proteins encompassing Der p 2 with either mature or proDer p 1 in Escherichia coli strain BL21 (DE3) and Origami2 (DE3), or in Pichia pastoris strain GS115 or X33. Four fusion proteins are expressed in Escherichia coli as inclusion bodies, whereas only chimeras comprising proDer p 1 are obtained in yeast
-
preparation described elsewhere, proform secreted in Pichia pastoris culture supernatant and converted into their prosequence-removed mature form, recombinant protein exhibits cysteine protease activity but no serine protease activity
-
recombinant ProDer p 1 from CHO cells
-
recombinant proforms secreted into culture supernatant of Pichia pastoris cells and converted into prosequence-removed mature form Der p 1-WT
-
the propiece of Der p 1 is PCR cloned and expressed in Escherichia coli
-
wild-type and mutant enzyme N52Q, expression in Pichia pastoris
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C104S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C118S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C32S/C72S/N63Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C32S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C35S
-
catalytic site mutant, expressed in Saccharomyces cerevisiae
C35S/N53Q
-
proform of mutant enzyme is efficiently secreted into culture medium. The secreted pro-C35S/N53Q is converted to the mature form by treatment in acidic conditions. Cysteine protease activity of the processed C35S/N53Q is lower than that of native Der f1 but significantly higher than that of the proenzyme
C4S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C66S/C104S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
C66S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
N53Q
-
the proform of N53Q is efficiently secreted into culture supernatant and processed to be of almost the same molecular weight as native enzyme by treatment in acidic conditions
N53Q
-
molecular weight identical with that of natural allergen
N52Q
-
mutant enzyme has no yeast-derived N-glycosylation
N52Q
-
mutant without glycosylation
N52Q
-
mutant with disrupted N-glycosylation motif in the prodomain, decreases the productivity and accelerates the maturation
N52Q
-
mutant with disrupted N-glycosylation motif in the prodomain, lower productivity
N52Q
-
mutant with disrupted N-glycosylation motif in the prodomain
N52Q
-
molecular weight identical with that of natural allergen
C72S/N53Q
-
proform of mutant enzyme is not efficiently secreted into culture medium
additional information
-
lysylendopeptidase-treated reDER f1 E(-1)K shows some differences in protease activity and reactivity with lectins: GlcNAc is contained at the non-reducing terminus of the sugar chains of native Der f1 but not of lysylendopeptidase-treated reDEr f1 E(-1)K. Lysylendopeptidase-treated reDEr f1 E(-1)K reacts with Lens culinaris agglutinin but native Der f1 does not, suggesting that fucose is connected with GlcNAc, bound to an asparagine residue, but not of native Der f1. Their N-terminal amino acid and the IgE-binding activity are the same as those of the native one
additional information
-
mutant Der f 1-N53Q, in which the N-glycosylation motif within the mature sequence is disrupted
C132V
-
site-directed mutagenesis, Der p 1 catalytic site mutant
additional information
-
hybrid molecules from parts of Der p 1 and Der p2 are constructed
additional information
-
development of chimeras assembling Der p 1 and Der p 2 allergens. Mutations in the Der p 1 catalytic site are also engineered
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ProDer p 1, refolding after solubilization in urea
-
recombinant His-tagged fusion proteins encompassing Der p 2 with either mature or proDer p 1 from Escherichia coli inclusion bodies with purification via denaturating nickel affinity chromatography
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
the efficient system to prepare active recombinant enzyme is useful for diagnosis and standardized allergen-specific immunotherapy for house dust mite allergy. The system would by a tool for analysis of IgE epitopes, determination of tertiary structure, allergen engineering for safer and more effective allergen-specific immunotherapy
medicine
-
Der f 1 is one of the most potent indoor allergens from house dust mites, shows IgE-eliciting activity in mice, recombinant DER f 1-N53Q will be a useful tool as alternatives to the natural Der f1 for in vitro and in vivo analyses
medicine
-
contribution of Der f 1 is particularly relevant in the mattresses and much less so in floor dust from the bedroom or living room, mattress covers (Goratex bedding system) seem to be an obvious option for lowering allergen exposure in sensitized individuals, geographical differences in prevelance of Dermatophagoides farinae in the Netherlands
medicine
-
cystatin A produced by keratinocytes is the dominant biochemical skin barrier against mite cysteine proteases
medicine
-
prodomains of Der f 1 reduce allergenicity, decreased activity of the proforms in inhibiting IgE binding, proforms exhibit different secondary structures than the mature forms, major conformational IgE epitopes commonly found in a broad population of patients exist within the 2 regions blocked by the prosegments, results basis for allergen standardization and design of safer and more effective allergen vaccines
medicine
-
upholstered seats in workplaces in Italy may constitue a significant reservoir of Der f1, significant correlation between indoor temperature and Der f 1 levels on floors, expressed per surface (no correlation when levels were expressed per weight), no correlation between concentration of mite allergen and geographical location, floor above ground, type of ventilation and indoor humidity, exposure to this allergen in workplaces may represent a risk factor for elicitation of symptoms and/or induction/maintenance of inflammation in allergic individuals and for sensitization
diagnostics
-
maturation systems using Pro-Der p 1 without the prodomain glycosylation are useful for the efficient preparation of a recombinant mature allergen
diagnostics
-
suspension-cultured BY-2 tobacco cells represent a low cost and environmentally safe expression system suitable to produce recombinant allergens from Dermatophagoides pteronyssinus under a form appropriate for diagnostic and therapeutic purposes
medicine
-
involved in the pathogenesis of allergy
medicine
-
proteolytic activity in pathogenesis of allergens
medicine
-
induces biological responses in a human airway-derived epithelial cell line
medicine
-
allergen is associated with allergic diseases such as bronchial asthma, rhinitis, and atopic dermatitis
medicine
-
affects individuals with allergy, resulting in rhinitis, asthma and/or atopic dermatitits
medicine
-
mimotopes identify conformational B-cell epitopes on the house dust mite allergen Der p 1. These mimotopes are potential candidates for the directed induction of blocking antibodies and epitope-specific immunotherapy of mite allergy
medicine
-
chimeras assembling Der p 1 and Der p 2 allergens are potential candidate vaccines against house dust mite allergy. But significant solubility and stability issues can limit the application of such chimeras for immunotherapy or diagnostic