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Information on EC 3.4.24.B16 - protease lasA and Organism(s) Pseudomonas aeruginosa and UniProt Accession P14789
for references in articles please use BRENDA:EC3.4.24.B16preliminary BRENDA-supplied EC number
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3.4.24.B16 protease lasASpecify your search results
Word Map
- 3.4.24.B16
- aeruginosa
- elastase
-
quorum
- pyocyanin
- lysostaphin
- rhamnolipids
- globisporus
-
pyoverdine
-
sensing-controlled
- pentaglycine
- lyticus
-
qs-controlled
- medicine
- degradation
The taxonomic range for the selected organisms is: Pseudomonas aeruginosa
The expected taxonomic range for this enzyme is: Pseudomonas aeruginosa
The expected taxonomic range for this enzyme is: Pseudomonas aeruginosa
Synonyms
bacteriolytic enzyme, lasa protease, staphylolysin, staphylolytic enzyme, protease lasa, lasa endopeptidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
staphylolysin
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
proteolytic degradation of proteins
substrate carbonyl oxygen displaces Zn2+-bound water molecule 2, thereby enabling direct interaction with Zn2+ to polarize the peptide carbonyl bond, rendering it susceptible to nucleophilic attack by the second Zn2+-bound water molecule, water molecule1. Water molecule 1 is oriented by interactions with the Nepsilon atoms of both His120 and His81, while that with Zn2+ is weakened. Either of these residues is capable of abstracting a proton from water molecule 1, enabling its addition, as hydroxide, to the substrate carbonyl carbon to generate an oxyanion that is stabilized by bidentate co-ordination of Zn2+ and by interaction with the unprotonated His residue. Proton transfer from the histidine general base to the departing amide nitrogen facilitates cleavage of the peptide bond to generate a product complex
proteolytic degradation of proteins
enzyme might be a modified serine protease with a His residue in the active site, specifically involved in degradation of elastin
-
proteolytic degradation of proteins
His120 is critical for LasA activity
-
proteolytic degradation of proteins
specific for Gly-Ala peptide bonds within Gly-Gly-Ala sequences in elastin, substrates that contain no Gly-Gly peptide bond, such as beta-casein, appear to be resistant to the enzyme
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CAS REGISTRY NUMBER
COMMENTARY
66554-76-5
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
4-(dimethylamino)azobenzene-4'-sulfonyl chloride-Leu-Gly-Gly-Gly-Ala-5-(2-aminoethylamino)-1-naphthalenesulfonic acid + H2O
?
-
-
-
-
?
beta-casein + H2O
beta-casein + 30 amino acid residues fragment
-
only 1 cleavage site: in the sequence NKKIEKFQphosphorylated-S between Lys and Ile, serine protease activity
-
?
succinyl-Gly-Gly-Leu-4-nitroanilide + H2O
Leu-4-nitroanilide + succinyl-Gly-Gly
-
-
-
-
?
succinyl-Gly-Gly-Phe-4-nitroanilide + H2O
Phe-4 nitroanilide + succinyl-Gly-Gly
-
better substrate than succinyl-Gly-Gly-Leu-4-nitroanilide
-
-
?
Elastin + H2O
?
-
cleavage of elastin by the enzyme possibly results in the unfolding or disruption of the complex elastin structure, thus providing more access to elastase and other proteases, elastolytic activity in absence of elastase, enzyme also enhances the elastolytic activity of elastase together with alkaline phophatase
-
?
Elastin + H2O
?
-
soluble recombinant and insoluble elastin
-
?
Elastin + H2O
?
-
contributes to the pathogenesis of Pseudomonas aeruginosa
-
?
Elastin + H2O
?
-
enzyme activates elastase, and the elastolytic activity of other proteases by cleaving elastin
-
?
additional information
?
-
-
enzyme activates elastase, and the elastolytic activity of thermolysin, human neutrophil elastase, proteinase K, by interacting with the elastin substrate rather than the other enzyme
-
?
additional information
?
-
-
no activity with tosyl-Gly-Pro-Lys-4-nitroanilide, enzyme activates elastase, and the elastolytic, not the proteolytic, activity of thermolysin, human neutrophil elastase, proteinase K, by interacting with the elastin substrate rather than the other enzymes
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells by cleaving the peptidoglycan pentaglycine interpeptides
-
?
additional information
?
-
-
no cleavage of beta-casein, does not bind chitin
-
-
?
additional information
?
-
-
no hydrolysis of 4-(dimethylamino)azobenzene-4'-sulfonyl chloride-Ala-Ala-Phe-Ala-5-(2-aminoethylamino)-1-naphthalenesulfonic acid
-
-
?
additional information
?
-
-
staphylolytic activity is pentaglycine in the peptidoglycan of Staphylococcus aureus
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
Elastin + H2O
?
-
contributes to the pathogenesis of Pseudomonas aeruginosa
-
?
Elastin + H2O
?
-
enzyme activates elastase, and the elastolytic activity of other proteases by cleaving elastin
-
?
additional information
?
-
-
enzyme activates elastase, and the elastolytic activity of thermolysin, human neutrophil elastase, proteinase K, by interacting with the elastin substrate rather than the other enzyme
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells
-
?
additional information
?
-
-
enzyme specifically promotes growth inhibition and lysis of Staphylococcus aureus cells by cleaving the peptidoglycan pentaglycine interpeptides
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Zinc
the active site of uncomplexed LasA contains a five-coordinate zinc ion with trigonal bipyramidal geometry and two metal-bound water molecules, crystallization data
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Nalpha-p-tosyl-L-lysine chloromethyl ketone
-
inhibition of the staphylolytic activity
additional information
-
no inhibition by 1,7-penanthroline and 4,7-phenanthroline, activity with beta-caseinis blocked by inhibitors that do not block the staphylolytic activity, no inhibition by several serine or cysteine proteinase inhibitors including diisopropyl fluorophosphate, phenylmethylsulfonylfluoride, leupeptin, and N-ethylmaleimide
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
elastase enhances staphylolytic activity of LasA
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.1
4-(dimethylamino)azobenzene-4'-sulfonyl-Leu-Gly-Gly-Gly-Ala-5-(2-aminoethylamino)-1-naphthalenesulfonic acid
-
in 5 mM Tris-HCl buffer, pH 8.5
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
15.2
4-(dimethylamino)azobenzene-4'-sulfonyl-Leu-Gly-Gly-Gly-Ala-5-(2-aminoethylamino)-1-naphthalenesulfonic acid
-
in 5 mM Tris-HCl buffer, pH 8.5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
active processed 20 kDa enzyme, member of the beta-lytic endopeptidase family of extracellular bacterial proteases
-
-
enzyme is synthesized in the cytoplasm as an inactive precursor which becomes active during translocation to the extracellular space
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
presence of Pseudomonas aeruginosa alters Staphylococcus aureus susceptibility to bactericidal antibiotics in a variable, strain-dependent manner. LasA endopeptidase potentiates lysis of Staphylococcus aureus by vancomycin. Vancomycin treatment of a Staphylococcus aureus mouse burn infection is potentiated by the presence of a LasA-producing Pseudomonas aeruginosa population
physiological function
-
LasA is a virulence determinant contributing to both acute and chronic Pseudomonas aeruginosa infections in immuno-compromised patients
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
19963
x * 19963, electrospray mass spectrometry, mature protein
20000
42000
45582
-
x * 20000, LasA, SDS-PAGE, x * 42000, proLasA, SDS-PAGE, x * 45582, pre-proLasA, DNA sequence determination
20000
-
x * 20000, LasA, SDS-PAGE, x * 42000, proLasA, SDS-PAGE, x * 45582, pre-proLasA, DNA sequence determination
42000
-
SDS-PAGE, LasA is synthesized as a preproenzyme that undergoes proteolysis to remove a 22000 Da amino-terminal propeptide
42000
-
x * 20000, LasA, SDS-PAGE, x * 42000, proLasA, SDS-PAGE, x * 45582, pre-proLasA, DNA sequence determination
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 20000, LasA, SDS-PAGE, x * 42000, proLasA, SDS-PAGE, x * 45582, pre-proLasA, DNA sequence determination
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
cleavage of proLasA precursor at position 237 of full-length gene product
proteolytic modification
proteolytic modification
-
enzyme precursor contains an elastase cleavage motif at the N-terminus, the enzyme is probably activated by elastase to activate the elastase afterwards
proteolytic modification
-
no autoproteolysis, enzyme is synthesized as a pre-proLasA, the signal sequence of pre-proLasA is cleaved off by a signal peptidase, the remaining inactive 42 kDa proLasA is secreted to the extracellular space, where the proform is cleaved to the mature active enzyme via a 28 kDa intermediate, plus a 14 kDa fragment, by elastase and a lysine specific protease, not by alkaline proteinase
proteolytic modification
-
no autoproteolysis, the 31 amino acid residues signal sequence of pre-proLasA is cleaved off by a signal peptidase, processing of the remaining 42 kDa proLasA by incubation with the culture filtrate of Pseudomonas aeruginosa or trypsin to the active 20 kDa LasA
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of active LasA as a complex with tartrate and in the uncomplexed form, to 1.28 A resolution. The overall fold resembles that of the other M23 family members, the LasA active site is less constricted and utilizes a different set of metal ligands. The active site of uncomplexed LasA contains a five-coordinate zinc ion with trigonal bipyramidal geometry and two metal-bound water molecules. Manual docking study of the pentapeptide Gly-Gly-Phe-Gly-Gly in the active site so that the carbonyl oxygen of the scissile peptide occupies the approximate position of Tyr151-bound tartrate oxygen O1 and the P2 and P1 glycines follow the path of the tartrate carbon skeleton
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H120A
-
oligonucleotide-directed mutagenesis, formation of the lasA5 allele, processing to a 20 kDa LasA-H120A protein and secretion to the medium occurs, but the mutant enzyme has no staphylolytic activity
H94A
-
mutation is moderately unstable and allows the production of a LasA protein with low enzymatic activity
I101A
-
mutation is moderately unstable and allows the production of a LasA protein with low enzymatic activity
L92A
-
mutation results in highly unstable protein that is susceptible to proteolytic degradation
N102A
-
mutation is moderately unstable and allows the production of a LasA protein with low enzymatic activity
W95A
-
mutation results in highly unstable protein that is susceptible to proteolytic degradation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis, overexpression of gene lasA in Escherichia coli strain JM109 in form of the pre-proLasA precursor protein
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression of LasA and LasB genes is decreased by Satureja khuzistanica essential oil extract. The MICs of the extract for different multidrug resistant isolates are 8, 8-12 microg/ml
-
the production of LasA is reduced by Terminalia catappa active extract (fraction 12) in a concentration dependent manner with about 50% reduction at 0.0625 mg/ml
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
degradation
-
two-stage enzymatic reaction for the continuous measurement of LasA protease activity using a defined substrate, supplemented with Streptomyces griseus aminopeptidase for further cleavage of the product, rate of release of the chromophore can be measured spectrophotometrically
medicine
additional information
-
cell lysis of Staphylococcus aureus, Micrococcus radiodurans and Gaffkia tetragena
medicine
staphylolysin is effective in the treatment of experimental methicillin-resistant Staphylococcus aureus-induced endophthalmitis in rats, and causes no morphological adverse effects to ocular tissues. Staphylolysin may be beneficial in the treatment of Staphylococcus aureus endophthalmitis in humans
medicine
presence of Pseudomonas aeruginosa alters Staphylococcus aureus susceptibility to bactericidal antibiotics in a variable, strain-dependent manner. LasA endopeptidase potentiates lysis of Staphylococcus aureus by vancomycin. Vancomycin treatment of a Staphylococcus aureus mouse burn infection is potentiated by the presence of a LasA-producing Pseudomonas aeruginosa population
medicine
-
LasA protease is effective in the treatment of experimental Staphylococcus aureus keratitis in rabbits, in both early- and late-treatment protocols the clinical scores for eyes treated with LasA protease are significantly lower than those for the eyes of corresponding controls
medicine
-
LasA protease is an effective therapeutic agent for Staphylococcus aureus experimental keratitis. The enzyme provides long-lasting protection against several strains of Staphylococcus aureus (MSSA, MRSA COL, MRSA 02/48406, and MRSA 02/48103) with IC50 values of 0.0018-0.005 mg/ml
medicine
-
the expression of LasA and LasB genes is decreased by Satureja khuzistanica essential oil extract. The MICs of the extract for different multidrug resistant isolates are 8-12 microg/ml
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Carnicero, A.; Mansito, T.B.; Roldan, J.M.; Falcon, M.A.
Staphylolytic enzyme from Pseudomonas aeruginosa: characterization and immunocytochemical localization
Arch. Microbiol.
154
37-41
1990
Pseudomonas aeruginosa
Peters, J.E.; Galloway, D.R.
Purification and characterization of an active fragment of the LasA protein from Pseudomonas aeruginosa: enhancement of elastase activity
J. Bacteriol.
172
2236-2240
1990
Pseudomonas aeruginosa
Gustin, J.K.; Kessler, E.; Ohman, D.E.
A substitution at His-120 in the LasA protease of Pseudomonas aeruginosa blocks enzymatic activity without affecting propeptide processing or extracellular secretion
J. Bacteriol.
178
6608-6617
1996
Pseudomonas aeruginosa, Pseudomonas aeruginosa FRD1
Kessler, E.; Safrin, M.; Abrams, W.R.; Rosenbloom, J.; Ohman, D.E.
Inhibitors and specificity of Pseudomonas aeruginosa LasA
J. Biol. Chem.
272
9884-9889
1997
Pseudomonas aeruginosa
Kessler, E.; Safrin, M.; Gustin, J.K.; Ohman, D.E.
Elastase and the LasA protease of Pseudomonas aeruginosa are secreted with their propeptides
J. Biol. Chem.
273
30225-30231
1998
Pseudomonas aeruginosa, Pseudomonas aeruginosa FRD740
Peters, J.E.; Park, S.J.; Darzins, A.; Freck, L.C.; Saulnier, J.M.; Wallach, J.M.; Galloway, D.R.
Further studies on Pseudomonas aeruginosa LasA: analysis of specificity
Mol. Microbiol.
6
1155-1162
1992
Pseudomonas aeruginosa
Kessler, E.; Safrin, M.; Blumberg, S.; Ohman, D.E.
A continuous spectrophotometric assay for Pseudomonas aeruginosa LasA protease (staphylolysin) using a two-stage enzymatic reaction
Anal. Biochem.
328
225-232
2004
Pseudomonas aeruginosa, Pseudomonas aeruginosa FRD2128(pJKGT107)
Barequet, I.S.; Ben Simon, G.J.; Safrin, M.; Ohman, D.E.; Kessler, E.
Pseudomonas aeruginosa LasA protease in treatment of experimental staphylococcal keratitis
Antimicrob. Agents Chemother.
48
1681-1687
2004
Pseudomonas aeruginosa, Pseudomonas aeruginosa FRD2128(pJKGT107)
Kessler, E.; Ohman, D.E.
Staphylolysin
Handbook of Proteolytic Enzymes(Barrett,A. J. ,Rawlings,N. D. ,Woessner,J. F. ,Eds. )Academic Press
1
1001-1003
2004
Pseudomonas aeruginosa
-
Elston, C.; Wallach, J.; Saulnier, J.
New continuous and specific fluorometric assays for Pseudomonas aeruginosa elastase and LasA protease
Anal. Biochem.
368
87-94
2007
Pseudomonas aeruginosa
Grande, K.K.; Gustin, J.K.; Kessler, E.; Ohman, D.E.
Identification of critical residues in the propeptide of LasA protease of Pseudomonas aeruginosa involved in the formation of a stable mature protease
J. Bacteriol.
189
3960-3968
2007
Pseudomonas aeruginosa
Tingpej, P.; Smith, L.; Rose, B.; Zhu, H.; Conibear, T.; Al Nassafi, K.; Manos, J.; Elkins, M.; Bye, P.; Willcox, M.; Bell, S.; Wainwright, C.; Harbour, C.
Phenotypic characterization of clonal and nonclonal Pseudomonas aeruginosa strains isolated from lungs of adults with cystic fibrosis
J. Clin. Microbiol.
45
1697-1704
2007
Pseudomonas aeruginosa
Barequet, I.S.; Habot-Wilner, Z.; Mann, O.; Safrin, M.; Ohman, D.E.; Kessler, E.; Rosner, M.
Evaluation of Pseudomonas aeruginosa staphylolysin (LasA protease) in the treatment of methicillin-resistant Staphylococcus aureus endophthalmitis in a rat model
Graefes Arch. Clin. Exp. Ophthalmol.
247
913-917
2009
Pseudomonas aeruginosa (P14789), Pseudomonas aeruginosa
Spencer, J.; Murphy, L.M.; Conners, R.; Sessions, R.B.; Gamblin, S.J.
Crystal structure of the LasA virulence factor from Pseudomonas aeruginosa: substrate specificity and mechanism of M23 metallopeptidases
J. Mol. Biol.
396
908-923
2010
Pseudomonas aeruginosa (P14789)
Barequet, I.S.; Bourla, N.; Pessach, Y.N.; Safrin, M.; Yankovich, D.; Ohman, D.E.; Rosner, M.; Kessler, E.
Staphylolysin is an effective therapeutic agent for Staphylococcus aureus experimental keratitis
Graefes Arch. Clin. Exp. Ophthalmol.
250
223-229
2012
Pseudomonas aeruginosa
Taganna, J.C.; Quanico, J.P.; Perono, R.M.; Amor, E.C.; Rivera, W.L.
Tannin-rich fraction from Terminalia catappa inhibits quorum sensing (QS) in Chromobacterium violaceum and the QS-controlled biofilm maturation and LasA staphylolytic activity in Pseudomonas aeruginosa
J. Ethnopharmacol.
134
865-871
2011
Pseudomonas aeruginosa, Pseudomonas aeruginosa ATCC 10145
Islamieh, D.I.; Afshar, D.; Esmaeili, D.
Effect of Satureja khuzistanica essential oil (SKEO) extract on expression of lasA and lasB genes in Pseudomonas aeruginosa
Iran. J. Microbiol.
11
55-59
2019
Pseudomonas aeruginosa
Radlinski, L.; Rowe, S.E.; Kartchner, L.B.; Maile, R.; Cairns, B.A.; Vitko, N.P.; Gode, C.J.; Lachiewicz, A.M.; Wolfgang, M.C.; Conlon, B.P.
Pseudomonas aeruginosa exoproducts determine antibiotic efficacy against Staphylococcus aureus
PLoS Biol.
15
e2003981
2017
Pseudomonas aeruginosa (P14789), Pseudomonas aeruginosa
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