Information on EC 3.4.21.81 - Streptogrisin B

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The expected taxonomic range for this enzyme is: Streptomyces griseus

EC NUMBER
COMMENTARY hide
3.4.21.81
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RECOMMENDED NAME
GeneOntology No.
Streptogrisin B
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of proteins with trypsin-like specificity
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
55071-87-9
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3 acetyl-Pro-Ala-Phe-Phe-NH2 + H2O
acetyl-Pro-Ala-Phe-Phe + NH3 + acetyl-Pro-Ala + Phe-Phe-NH2 + acetyl-Pro-Ala-Phe + Phe-NH2
show the reaction diagram
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hydrolyzed at the Ala-Phe, the Phe-Phe and the Phe-NH2 bond
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-
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acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide + H2O
acetyl-Ala-Ala-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
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-
-
-
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Acetyl-Pro-Ala-Ala-Phe-NH2 + H2O
?
show the reaction diagram
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split at the amide bond as well as at the Ala-Ala bond
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-
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Acetyl-Pro-Ala-Gly-Phe-NH2 + H2O
?
show the reaction diagram
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split at the amide bond as well as at the Ala-Gly bond
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-
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Acetyl-Pro-Ala-Leu-Phe-NH2 + H2O
Acetyl-Pro-Ala-Leu-Phe-OH + NH3
show the reaction diagram
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hydrolyzed at the amide bond only
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-
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Acetyl-Pro-Ala-Pro-Phe-NH2 + H2O
Acetyl-Pro-Ala-Pro-Phe-OH + NH3
show the reaction diagram
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hydrolyzed at the amide bond only
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-
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N-Acetyl-L-Pro-L-Leu-p-nitroanilide + H2O
?
show the reaction diagram
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-
-
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OMSVP3 + H2O
?
show the reaction diagram
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ovomucoid third domain from silver pheasant, potent inhibitor of chymotrypsin, subtilisin, and elastase
cleavage between residues Met18-Glu19
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?
OMSVP3 mutant P14C/N39C + H2O
?
show the reaction diagram
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disulfide variant of the ovomucoid third domain from silver pheasant carrying an engineered Cys14-Cys39 bond near the reactive site
cleavage between residues Met18-Glu19. P14C/N39C can be selectively cleaved by Streptomyces griseus protease B at the reactive site of OMSVP3 to form a reactive site modified inhibitor. The conversion rate of P14C/N39C is much faster than that for wild type under any pH condition. The reactive site modified form of P14C/N39C is thermodynamically more stable than the intact one
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?
Sc-AAPF-p-nitroanilide + H2O
?
show the reaction diagram
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-
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?
Sc-AAPF-SBn + FAASF-NH2
?
show the reaction diagram
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ligation to nonapeptide by mutant S195A/T213L/F228H
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?
Sc-AAPF-SBn + H2O
?
show the reaction diagram
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-
-
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Suc-AAPF-p-nitroanilide + H2O
?
show the reaction diagram
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-
-
?
Suc-AAPF-SBzl + H2O
Suc-AAPF + phenylmethanethiol
show the reaction diagram
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-
-
?
succinyl-Ala-Ala-Pro-4-methylcoumarin 7-amide + H2O
succinyl-Ala-Ala-Pro + 7-amino-4-methylcoumarin
show the reaction diagram
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succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
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-
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succinyl-Ala-Ala-Pro-X-4-nitroanilide + H2O
?
show the reaction diagram
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-
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?
succinyl-Ala-Ala-Pro-X-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
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?
succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Asp + 4-nitroaniline
show the reaction diagram
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extremely poor substrate at pH 8 but becomes a moderately good substrate at pH 5.5
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-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Leu + 4-nitroaniline
show the reaction diagram
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optimum at pH 7.5-8.0
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?
succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Lys + 4-nitroaniline
show the reaction diagram
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optimum at pH 10.0
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?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Bovine pancreatic trypsin inhibitor
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Kunitz
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N-tert-Butyloxycarbonyl-glycyl-L-leucyl-L-phenylalanine chloromethyl ketone
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N-tert-Butyloxycarbonyl-L-alanylglycyl-L-phenylalanine chloromethyl ketone
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protein inhibitor S13D14Y15-OMTKY3
a OMTKY3 mutant variant, recombinantly produced in Escherichia coli strain RV308 and purified, strong inhibition, overview
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protein inhibitor S13D14Y15G18I19K21-OMTKY3
a OMTKY3 mutant variant, recombinantly produced in Escherichia coli strain RV308 and purified, strong inhibition, overview. S13D14Y15G18I19K21 OMTKY3 inhibits Streptomyces griseus protease B 10times more strongly than Streptomyces griseus protease A, EC 3.4.21.80
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Soybean trypsin inhibitor
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Squash inhibitor
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Streptomyces subtilisin inhibitor
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Third domain of the ovomucoid inhibitor from turkey
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Turkey ovomucoid third domain
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OMTKY3, inhibitor for serine peptidases
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
Acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide
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1.4
Acetyl-Pro-Ala-Leu-Phe-NH2
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2.4
Acetyl-Pro-Ala-Pro-Phe-NH2
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0.062 - 0.5
Suc-AAPF-p-nitroanilide
0.101
succinyl-Ala-Ala-Pro-Phe 4--nitroanilide
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0.098
Succinyl-Ala-Ala-Pro-Phe-4-methylcoumarin 7-amide
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additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04
Acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide
Streptomyces griseus
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7.1
Acetyl-Pro-Ala-Leu-Phe-NH2
Streptomyces griseus
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6.7
Acetyl-Pro-Ala-Pro-Phe-NH2
Streptomyces griseus
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0.066 - 1884
Suc-AAPF-p-nitroanilide
153
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
Streptomyces griseus
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3.3
Succinyl-Ala-Ala-Pro-Phe-4-methylcoumarin 7-amide
Streptomyces griseus
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additional information
additional information
Streptomyces griseus
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
740 - 10700
succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide
168017
28200 - 25000000
succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide
168018
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide
6
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hydrolysis reaction
7.5 - 8
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide
8.8 - 9.2
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ligation reaction of mutant S195A/T213L/F228H
10
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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ratio of ligation to hydrolysis by mutant S195A/T213L/F228H decreases with increase in temperature from 30°C to 60°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
18410
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Streptomyces griseus, amino acid sequence
18412
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1 * 18412, Streptomyces griseus, amino acid sequence
additional information
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amino acid sequence of cysteic acid peptides from peptic digest
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
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enzyme folds to native state that is thermodynamically marginally stable, apparent stability arises kinetically from unfolding free energy barriers. Enzyme has effective pro-regions to facilitate folding
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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enzyme has effective pro-regions to facilitate folding
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
bound to wild type OMTKY3 and to the Ala32I variant, by the hanging-drop vapor-diffusion method, SGPB and OMTKY3 Ala32I forms a 1 : 2 complex in the crystal
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crystal structure in complex with turkey ovomucoid thir domain backbone variants OMTKY3-Pro18I and OMTKY3-psi[COO]-Leu18I
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crystal structure of protease B with tripeptide chloromethyl ketone inhibitors
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crystal structures of Leu18, Ala18, and Gly18 variants of turkey ovomucoid inhibitor third domain complexed with proteinase B
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structure at 2.8 A resolution
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structure of the complex of protease B with the third domain of the ovomucoid inhibitor from turkey at 1.8 A resolution
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structure of the complex of proteinase B and polypeptide chymotrypsin inhibitor-1 from Russet Burbank potato tubers at 2.1 A resolution
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native crystal structures at pH 4.2 at a resolution of 1.18 A, and at pH 7.3 at a resolution of 1.23 A. At pH 4.2 the enzyme is assigned as a tetrapeptide, Asp-Ala-Ile-Tyr, whereas at pH 7.3 it is assigned as a tyrosine molecule and a leucine molecule existing at equal occupancies in both of the SGPB molecules in the asymmetric unit
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 45
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mutant S195A, completely inactivated in less than 1 min at 45°C, less than 20 min at 37°C
additional information
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study of temperature-dependences of the folding and unfolding kinetics without enzyme pro region
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Stable in presence of 6.0 M guanidinium chloride
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
guanidine-HCl
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8 M, extremely stable
urea
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6 M, extremely stable
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
further purification of a commercial preparation
mutants S195A and S195G
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purified from a commercial preparation of Pronase
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purified from Pronase
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
active-site variant genes clones in the Escherichia coli/Bacillus subtilis shuttle vector pEB-11 and expressed in Bacillus subtilis DB104
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gene structure
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genes constructed which encodes deletions at the amino-terminal end of the propeptide, deletions to the 76-amino-acid propeptide of SGPB cause an incremental loss in the production of the mature enzyme
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S195A
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active-site variant of protease B with peptide-ligation activity
S195A/T213L/F228H
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i.e. streptoligase, catalyzes peptide ligation efficiently. Ligation proceeds via an acyl-enzyme intermediate involving H57. Mutant exhibits half-life for unfolding of 16.3 min at 55°C in the absence of stabilizing substrates
S195C
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active-site variant of protease B with peptide-ligation activity
S195G
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active-site variant of protease B with peptide-ligation activity
S195G/H57N
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active-site variant of protease B with peptide-ligation activity
additional information
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introduction of site-specific disulfide bonds into the flexible N-terminal loop of natural Kazal-type inhibitors to characterize the thermodynamics of the reactive site peptide bond hydrolysis. Mutant P14C/N39C is a disulfide variant of the ovomucoid third domain from silver pheasant carrying an engineered Cys14–Cys39 bond near the reactive site. The conversion rate of P14C/N39C by streptogrisin B is much faster than that for wild type under any pH condition
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
study of temperature-dependencies of the folding and unfolding kinetics without enzyme pro region. Enzyme shows a maximal unfolding cooperativity and slow rate of global unfolding
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