Information on EC 3.4.22.46 - L-peptidase

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: Foot-and-mouth disease virus

EC NUMBER
COMMENTARY
3.4.22.46
-
RECOMMENDED NAME
GeneOntology No.
L-peptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
autocatalytically cleaves itself from the polyprotein of the foot-and-mouth disease virus by hydrolysis of a Lys-/-Gly bond, but then cleaves host cell initiation factor eIF-4G at bonds -Gly-/-Arg- and -Lys-/-Arg-
show the reaction diagram
Best known from foot-and-mouth disease virus, but occurs in other aphthoviruses and cardioviruses. Destruction of initiation factor eIF-4G has the effect of shutting off host-cell protein synthesis while allowing synthesis of viral proteins to continue. The tertiary structure reveals a distant relationship to papain and, consistent with this, compound E-64 is inhibitory. Type example of peptidase family C28
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Eukaryotic signal peptidase
-
-
-
-
Eukaryotic signal proteinase
-
-
-
-
foot-and-mouth disease virus 3C protease
-
-
L proteinase
-
-
-
-
Leader peptidase
-
-
-
-
Leader peptide hydrolase
-
-
-
-
Leader proteinase
-
-
-
-
Leader proteinase
Foot-and-mouth disease virus FMDV
-
-
-
Lpro
-
-
-
-
Lpro
Foot-and-mouth disease virus FMDV
-
-
-
Peptidase, signal
-
-
-
-
Prokaryotic leader peptidase
-
-
-
-
Prokaryotic signal peptidase
-
-
-
-
Prokaryotic signal proteinase
-
-
-
-
Propeptidase
-
-
-
-
Proteinase, eukaryotic signal
-
-
-
-
Proteinase, signal
-
-
-
-
Signal peptidase
-
-
-
-
Signalase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
934238-57-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
FMDV O1Kaufbeuren B64, from BHK or A293 cells, two enzymes: leader protease and 3C protease
-
-
Manually annotated by BRENDA team
FMDV, two forms of the leader proteinase, Lab, starting at the first AUG, and Lb, starting at second AUG
-
-
Manually annotated by BRENDA team
Foot-and-mouth disease virus FMDV
FMDV
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
significant conformational adaptation by the enzyme is important for substrate recognition, specificity differences between 3Cpro from different picornaviruses, overview
physiological function
-
the viral RNA genome is translated in the cytoplasm of infected cells as a long polyprotein precursor that is cleaved by virally encoded proteases to release functional proteins needed for the synthesis of new virions. In 10 of 13 cases, 3Cpro performs the cleavages by targeting specific sequences within the polyprotein
physiological function
-
the leader proteinase, Lpro, acts as an interferon-beta antagonist and inhibits IFN-alpha1/beta promoter activation, poly(I:C)-induced IFN-alpha1/beta mRNA expression, and dsRNA-induced type I interferon transcription by decreasing interferon regulatory factor 3/7 in protein levels. Lpro significantly reduced the transcription of multiple IRF-responsive genes
physiological function
-
foot and mouth disease virus expresses its genetic information as a single polyprotein that is translated from the single-stranded RNA genome. The leader protease, Lbpro, performs the initial cleavage by freeing itself from the growing polypeptide chain. Subsequently, Lbpro cleaves the two homologues of the host cell protein eukaryotic initiation factor 4G, eIF4G. Lbpro possesses specific binding sites at the non-prime side from S1 down to S7
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Abz-KLKGAGQ-EDDnp + H2O
Abz-KLK + GAGQ-EDDnp
show the reaction diagram
-
hydrolysis of FRET peptides
-
-
?
eukaryotic initiation factor 4G + H2O
?
show the reaction diagram
-
Lbpro cleaves two homologues of the host cell protein. Lbpro possesses specific binding sites at the non-prime side from S1 down to S7
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, recognition/cleavage site is ANLG*RTTL
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage between residues G674 and R675, the enzyme binds to substrate residues 640-669 and interacts with C133 and residues 183-195, interaction analysis of enzyme with recombinant peptide fragments, reduced binding with mutated peptides K643A, K646A, and R650A overview
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, recognition/cleavage site is ANLG*RTTL
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site, eukaryotic host cell protein substrate, cleavage between residues G674 and R675, the enzyme binds to substrate residues 640-669 and interacts with C133 and residues 183-195, interaction analysis of enzyme with recombinant peptide fragments, reduced binding with mutated peptides K643A, K646A, and R650A overview
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, recognition/cleavage site is LNVG*SRRS, but not ADFG*RQTP
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, recognition/cleavage site is LNVG*SRRS, but not ADFG*RQTP
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site, eukaryotic host cell protein substrate
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protein synthesis
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protien synthesis
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, activity of 3Cpro and Lpro with wild-type and mutant substrate, overview, the highly specific 3Cpro cleavage site is located at the small 40-amino-acid region of the protein
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protein synthesis, eukaryotic host cell protein substrate
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
-
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
cis and trans cleavage activity at the L/P1 junction
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
similar to other papain-like proteinases the enzyme possesses the catalytic cysteine and histidine residues, however the catalytic asparagine has been replaced by an aspartate. The interaction between the histidine and aspartate is exposed to solvent, as the tryptophan residues are missing. The cleavage site is between its own C-terminus and the N-terminus of VP4
-
-
?
human cyclin A + H2O
?
show the reaction diagram
-
-
-
-
?
Val-Gln-Arg-Lys-Leu-Lys-4-methylcoumarin 7-amide + H2O
Val-Gln-Arg-Lys-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
the substrate corresponds to the six C-terminal amino acids of the leader protein
-
?
mengovirus polypeptide + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
no hydrolysis of Phe-Arg-4-methylcoumarin 7-amide, Val-Lue-Lys-4-methylcoumarin 7-amide, classical papain substrates, or Ser-Phe-Ala-Asn-Leu-Gly-4-methylcoumarin 7-amide, corresponding to the N-terminal portion of the eIF4G cleavage site
-
-
-
additional information
?
-
-
the enzyme inhibits the immediate-early induction of beta interferon mRNA and blocks the host innate immune response by inhibition of IFN-induction of double-stranded RNA-dependent protein kinase R, 2',5'-oligoadenylate synthase, and Mx1 mRNAs in host porcine cells
-
-
-
additional information
?
-
-
elIF4GI cleavage site mapping
-
-
-
additional information
?
-
-
substrate recognition and cleavage site specificity, overview
-
-
-
additional information
?
-
-
the enzyme is synthesized as part of a large polyprotein LbproVP4VP, from which it releases itself by highly efficient self-processing between its C- and N-terminus of the subsequent protein VP4, recognition of the sequence QRKLK*GAGQ, specificity at the P2, Leu, and P3 positions, including S3 subsite preferring Lys or Asn, and comparison with other papain-like enzymes, no activity when Pro is at P2 position and with type I collagen, overview, residues P99, P100, L143, A149, and L178 determine the specificity of the S2 binding pocket
-
-
-
additional information
?
-
-
the leader protease, Lbpro, performs the initial cleavage by freeing itself from the growing polypeptide chain. Lbpro is not only one of the smallest papain-like cysteine peptidases but also one of the most specific. It has high prime side specificity at least down to the S' 5 site. It can still however cleave between both K-G and G-R pairs
-
-
-
additional information
?
-
-
the enzyme typically recognise the alternating positions of the side chains along the polypeptide sequence that are characteristic of an extended backbone conformation, and this serves to place the scissile bond in the correct orientation at the active site. Significant conformational adaptation by the enzyme is important for substrate recognition
-
-
-
additional information
?
-
Foot-and-mouth disease virus FMDV
-
the enzyme is synthesized as part of a large polyprotein LbproVP4VP, from which it releases itself by highly efficient self-processing between its C- and N-terminus of the subsequent protein VP4, recognition of the sequence QRKLK*GAGQ, specificity at the P2, Leu, and P3 positions, including S3 subsite preferring Lys or Asn, and comparison with other papain-like enzymes, no activity when Pro is at P2 position and with type I collagen, overview, residues P99, P100, L143, A149, and L178 determine the specificity of the S2 binding pocket
-
-
-
additional information
?
-
Foot-and-mouth disease virus FMDV
-
substrate recognition and cleavage site specificity, overview
-
-
-
additional information
?
-
Foot-and-mouth disease virus FMDV
-
the enzyme inhibits the immediate-early induction of beta interferon mRNA and blocks the host innate immune response by inhibition of IFN-induction of double-stranded RNA-dependent protein kinase R, 2',5'-oligoadenylate synthase, and Mx1 mRNAs in host porcine cells
-
-
-
poliovirus replicase-related polypeptide + H2O
?
show the reaction diagram
-
-
-
-
?
translation initiation factor eIF4G + H2O
additional information
-
-
-
-
-
?
translation initiation factor eIF4G + H2O
additional information
-
-
-
characterization of cleavage products, primary cleavage of rabbit reticulocyte eIF-4-gamma occurs between Gly479-Arg480, complete proteolysis after 12 h
?
translation initiation factor eIF4G + H2O
additional information
-
-
very rapid reaction
-
-
?
translation initiation factor eIF4G + H2O
additional information
-
-
i.e. p220
-
-
?
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
eukaryotic initiation factor 4G + H2O
?
show the reaction diagram
-
Lbpro cleaves two homologues of the host cell protein. Lbpro possesses specific binding sites at the non-prime side from S1 down to S7
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus, Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, recognition/cleavage site is ANLG*RTTL
-
-
?
eukaryotic initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
Foot-and-mouth disease virus, Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, recognition/cleavage site is LNVG*SRRS, but not ADFG*RQTP
-
-
?
eukaryotic initiation factor eIF4GII + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage prevents the synthesis of host cellular protein from capped cellular mRNAs, while the viral RNA is tsill translated initiating from an internal ribosome entry site
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protein synthesis
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protien synthesis
-
-
?
eukaryotic translation initiation factor eIF4GI + H2O
?
show the reaction diagram
Foot-and-mouth disease virus FMDV
-
eukaryotic host cell protein substrate, cleavage causes the rapid inhibition of cellular cap-dependent protein synthesis
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
-
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
cis and trans cleavage activity at the L/P1 junction
-
-
?
foot-and-mouth disease leader protein + H2O
?
show the reaction diagram
-
similar to other papain-like proteinases the enzyme possesses the catalytic cysteine and histidine residues, however the catalytic asparagine has been replaced by an aspartate. The interaction between the histidine and aspartate is exposed to solvent, as the tryptophan residues are missing. The cleavage site is between its own C-terminus and the N-terminus of VP4
-
-
?
additional information
?
-
-
the enzyme inhibits the immediate-early induction of beta interferon mRNA and blocks the host innate immune response by inhibition of IFN-induction of double-stranded RNA-dependent protein kinase R, 2',5'-oligoadenylate synthase, and Mx1 mRNAs in host porcine cells
-
-
-
additional information
?
-
-
the leader protease, Lbpro, performs the initial cleavage by freeing itself from the growing polypeptide chain. Lbpro is not only one of the smallest papain-like cysteine peptidases but also one of the most specific. It has high prime side specificity at least down to the S' 5 site. It can still however cleave between both K-G and G-R pairs
-
-
-
additional information
?
-
Foot-and-mouth disease virus FMDV
-
the enzyme inhibits the immediate-early induction of beta interferon mRNA and blocks the host innate immune response by inhibition of IFN-induction of double-stranded RNA-dependent protein kinase R, 2',5'-oligoadenylate synthase, and Mx1 mRNAs in host porcine cells
-
-
-
translation initiation factor eIF4G + H2O
additional information
-
-
-
-
-
?
translation initiation factor eIF4G + H2O
additional information
-
-
-
characterization of cleavage products, primary cleavage of rabbit reticulocyte eIF-4-gamma occurs between Gly479-Arg480, complete proteolysis after 12 h
?
translation initiation factor eIF4G + H2O
additional information
-
-
very rapid reaction
-
-
?
translation initiation factor eIF4G + H2O
additional information
-
-
i.e. p220
-
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
CA074
-
epoxide-based inhibitor of cysteine proteinases, two-step overall irreversible inhibition, with step one being reversible and step two irreversible
E64
-
epoxide-based inhibitor of cysteine proteinases, two-step overall irreversible inhibition, with step one being reversible and step two irreversible. Extending E64 by addition of the dipeptide R-P to a compound termed E64-R-P-NH2, which irreversibly inhibits Lbpro with a Ki of 30 nM and k4 of 0.01/min, can serve as the basis for design of specific inhibitors of FMDV replication
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
i.e. E-64
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
-
E64-R-P-NH2
-
extending E64 by addition of the dipeptide R-P to a compound termed E64-R-P-NH2, which irreversibly inhibits Lbpro with a Ki of 30 nM and k4 of 0.01/min, can serve as the basis for design of specific inhibitors of FMDV replication
-
additional information
-
sensitive to high ionic strength, 50% inactivation by 20 mM NaCl or 10 mM CaCl2
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0227
-
Val-Gln-Arg-Lys-Leu-Lys-4-methylcoumarin 7-amide
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0116
-
CA074
-
pH 7.8, 37C
0.0034
-
E64
-
pH 7.8, 37C
0.00003
-
E64-R-P-NH2
-
pH 7.8, 37C
-
additional information
-
additional information
-
kinetics mechanism of two-step overall irreversible inhibition by cysteinase inhibitors
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
self-processing activity of the enzyme on wild-type and mutant polyproteins, overview
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
-
substrate Val-Gln-Arg-Lys-Leu-Lys-4-methylcoumarin 7-amide
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
9.8
-
less than 50% of maximal activity above and below, substrate Val-Gln-Arg-Lys-Leu-Lys-4-methylcoumarin 7-amide
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
additional information
-
the L signal is mainly localized to the nuclei of infected cells, while VP1 is detected in the cytoplasm, thereafter the L signal is detected throughout the whole cell. The pattern of L sub-cellular localization of the L mutant viruses is similar to the WT virus, except for a delay in the cytoplasmic L protein signal
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
Lpro is localized to the nucleus of infected cells, and there is a correlation between the translocation of Lpro and the decrease in the amount of nuclear p65/RelA, a subunit of NF-kappa B
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Foot-and-mouth disease virus (isolate Bovine/Germany/O1Kaufbeuren/1966 serotype O)
Foot-and-mouth disease virus (isolate Bovine/Germany/O1Kaufbeuren/1966 serotype O)
Foot-and-mouth disease virus (isolate Bovine/Germany/O1Kaufbeuren/1966 serotype O)
Foot-and-mouth disease virus (isolate Bovine/Germany/O1Kaufbeuren/1966 serotype O)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20000
-
-
structure prediction
20000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
dimer in solution, determined by NMR-spectroscopy. Dimer can not be dissociated by increasing the ionic strength or by dilution
additional information
-
the peptide binding cleft, which contains the active site at its centre, is located at the interface between two beta-barrels. Unusually, picornaviral 3Cpro possess a Cys-His-Asp/Glu catalytic triad at the centre of this cleft, instead of the Ser-His-Asp arrangement of active-site residues
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the enzyme is synthesized as part of a large polyprotein LbproVP4VP2, from which it releases itself by highly efficient self-processing between its C- and N-terminus of the subsequent protein VP4, recognition of the sequence QRKLK*GAGQ, Asp at P3 and Phe at P2 severely compromise the self-processing
proteolytic modification
Foot-and-mouth disease virus FMDV
-
the enzyme is synthesized as part of a large polyprotein LbproVP4VP2, from which it releases itself by highly efficient self-processing between its C- and N-terminus of the subsequent protein VP4, recognition of the sequence QRKLK*GAGQ, Asp at P3 and Phe at P2 severely compromise the self-processing
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structures of LbproC51A, LbproC51A/C133S, sLbproC51A/C133S
-
foot-and-mouth disease virus 3C protease mutant C95K/C142L/C163A complexed with a peptide substrates APAKQLLNFD and APAKELLNFD that spans P5-P5', 1:5.5 molar ratio of enzyme to peptide, 0.05 ml of of 3Cpro solution at 17 mg/ml protein in 100 mM HEPES, pH 7.0, 400 mM NaCl, 1 mM EDTA, 2 mM 2-mercaptoethanol, and 0.01% w/v sodium azide is mixed with 0.007 ml of 30 mM peptide dissolved in the same buffer, yielding a final protein concentration of 14.9 mg/ml, sitting-drop vapour diffusion, room temperature, mixing of 0.001 ml of protein complex solution with 0.003 ml reservoir composed of 40-43% PEG 400, 0.2 M LiSO4 and 0.1 M Tris, pH 8.0 X-ray diffraction structure determination and analysis
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, phosphate buffer, 1 mM DTT, 50% glycerol
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant enzyme mutants from Escherichia coli by gel filtration
-
recombinant GST-tagged enzyme from Escherichia coli strain JM101 by glutathione affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli BL-21
-
expression of enzyme mutants in Escherichia coli
-
expression of FLAG- and HA-tagged FMDV A24-L1123 mutant enzyme in LF-BK cells. T7 RNA transcripts of SpeI-digested pA24-WT, pA24-L1123 and pA24-L tag mutants, as well as transcript of control plasmid pL, are translated in vitro in rabbit reticulocyte lysates. All synthesized L mutant proteins are able to cleave themselves from the precursor L-Vp4-Vp2', the Lbeta band is absent in all L mutant translation reactions. The tn insertion in these mutant L plasmids forces translation initiation in vitro from other than the second AUG codon
-
expression of GST-tagged enzyme in Escherichia coli strain JM101
-
expression of wild-type and mutant enzymes by in vitro transcription and translation using rabbit reticulocyte lysate
-
PK-15 cells are transfected with a HA-tagged Labpro expression construct, together with a luciferase reporter plasmid with the porcine IFN-alpha1 promoter or the IFN-beta promoter and pRL-TK. At 24 h post-transfection, the cells were further transfected or mock-transfected with poly(I:C) followed by the dual-luciferase assay, quantitative real-time RT-PCR, overview
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C95K/C142L/C163A
-
the C-terminally truncated, catalytically inactive mutant has wild-type binding activity but remains soluble at purified protein concentrations in excess of 10 mg/ml
C95K/C142S/C163A
-
C-terminally truncated, catalytically inactive form of the type A10FMDV 3Cpro
D136N
-
reduced activity
E48Q
-
active mutant
H110L
-
active mutant, nearly as active as wild-type
H120L
-
no activity
H81L
-
active mutant
L143A
P03305
self-processes efficiently at the L(pro)/VP4 cleavage site containing P2 phenylalanine, self-processing at the eIF4GII sequence Asp-Phe-Gly-Arg-Gln-Thr is improved but shows more-extensive aberrant processing
L143M
P03305
does not self-process efficiently at the L(pro)/VP4 cleavage site containing P2 phenylalanine
additional information
-
mutation of the viral polyprotein self-processing cleavage site QRKLK*GAGQ, replacement by several variants of the two protein substrate elIF4G cleavage sites ANLG*RTTL and LNVG*SRRS, overview
additional information
-
a mutant virus A12-LLV2 lacking the leader proteinase is attenuated in cell culture and susceptible animals due to the inability of the mutant virus to block the expression of type I interferon alpha/beta, the blockade results in the IFN-induced inhibition of FMDV replication, overview
additional information
-
Lpro regulates the activity of nuclear factor kappa B. Analysis of NF-kappa B-dependent reporter gene expression in BHK-21 cells (baby hamster kidney cells strain 21), demonstrate that infection with wild-type virus has an inhibitory effect compared to infection with a genetically engineered mutant (A12-LLV2) lacking the leader coding region
additional information
P03305
leader proteinase self-processes inefficiently at the L(pro)/VP4 cleavage site Lys-Leu-Lys-Gly-Ala-Gly when the leucine at position P2 is replaced by phenylalanine: Molecular modeling and energy minimization identifies the L(pro) residue L143 as being responsible for this discrimination
additional information
-
introduction of FLAG- and HA-tag on the small tetracysteine tc motif CCGPCC of cDNA-derived mutant FMDV A24-L1123, the mutant shows a small plaque phenotype similar to the parental A24-L1123 mutant. Expression of the Flag- or tc-tagged Lab protein is abolished or greatly diminished in these viruses. The A24-L1123/Flag virus acquires an extra base in the inter-AUG region that results in new AUG codons in-frame with the second AUG, and produced a larger Lb protein. This N-terminal extension of the Lb protein in mutant A24-L1123/Flag does not affect virus viability or L functions in cell culture
additional information
Foot-and-mouth disease virus FMDV
-
a mutant virus A12-LLV2 lacking the leader proteinase is attenuated in cell culture and susceptible animals due to the inability of the mutant virus to block the expression of type I interferon alpha/beta, the blockade results in the IFN-induced inhibition of FMDV replication, overview; mutation of the viral polyprotein self-processing cleavage site QRKLK*GAGQ, replacement by several variants of the two protein substrate elIF4G cleavage sites ANLG*RTTL and LNVG*SRRS, overview
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
drug development
-
the enzym eis a target for inhibitor design for inhibition of viral replication