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Literature summary for 3.4.21.62 extracted from

  • Howell, M.; Dumitrescu, D.G.; Blankenship, L.R.; Herkert, D.; Hatzios, S.K.
    Functional characterization of a subtilisin-like serine protease from Vibrio cholerae (2019), J. Biol. Chem., 294, 9888-9900 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.21.62

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of C-terminally His6- and FLAG-tagged wild-type and mutant enzymes in Escherichia coli Vibrio cholerae serotype O1

Protein Variants

Protein Variants Comment Organism
S361A site-directed mutagenesis, mutation of the catalytic residue partially inhibits autoproteolysis, but the IvaPS361A mutant can be cleaved to the 47-kDa mature form through a mechanism other than autoproteolysis Vibrio cholerae serotype O1

Inhibitors

Inhibitors Comment Organism Structure
IvaP I9 domain the N-terminal propeptide of enzyme IvaP, the IvaP I9 domain, can temporarily inhibit, and be cleaved by, the purified enzyme Vibrio cholerae serotype O1
additional information no inhibition by EDTA and benzamidine Vibrio cholerae serotype O1
PMSF strong inhibition Vibrio cholerae serotype O1

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular the enzyme proform contains a signal peptide and is secreted Vibrio cholerae serotype O1
-
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
intelectin + H2O Vibrio cholerae serotype O1 enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae ?
-
 ?
intelectin + H2O Vibrio cholerae serotype O1 C6706 enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae ?
-
 ?
intelectin + H2O Vibrio cholerae serotype O1 El Tor Inaba N16961 enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae ?
-
 ?
intelectin + H2O Vibrio cholerae serotype O1 ATCC 39315 enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae ?
-
 ?
IvaP I9 domain + H2O Vibrio cholerae serotype O1 a temporary inhibitor and substrate of purified IvaP ?
-
 ?
additional information Vibrio cholerae serotype O1 Ser361 contributes to IvaP autoprocessing in stationary-phase cultures ?
-
 ?
additional information Vibrio cholerae serotype O1 C6706 Ser361 contributes to IvaP autoprocessing in stationary-phase cultures ?
-
 ?
additional information Vibrio cholerae serotype O1 El Tor Inaba N16961 Ser361 contributes to IvaP autoprocessing in stationary-phase cultures ?
-
 ?
additional information Vibrio cholerae serotype O1 ATCC 39315 Ser361 contributes to IvaP autoprocessing in stationary-phase cultures ?
-
 ?

Organism

Organism UniProt Comment Textmining
Vibrio cholerae serotype O1 Q9KVI8
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-
Vibrio cholerae serotype O1 ATCC 39315 Q9KVI8
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-
Vibrio cholerae serotype O1 C6706 Q9KVI8
-
-
Vibrio cholerae serotype O1 El Tor Inaba N16961 Q9KVI8
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification IvaP undergoes extensive post-translational processing. Following secretion, enzyme IvaP is cleaved at least three times to yield a truncated enzyme with serine hydrolase activity, extracellular maturation requires a series of sequential N- and C-terminal cleavage events congruent with the enzyme's mosaic protein domain structure. IvaP can be partially processed in trans, but intramolecular proteolysis is most likely required to generate the mature enzyme. Unlike many other subtilisin-like enzymes, the IvaP cleavage pattern is consistent with stepwise processing of the N-terminal propeptide, which can temporarily inhibit, and be cleaved by, the purified enzyme. IvaP processing results in the loss of about 139 amino acids (about 15 kDa) from the IvaP N-terminus and about 23 amino acids (about 3 kDa) from the IvaP C-terminus, cleavage pattern overview. These sequencing results are consistent with N-terminal cleavage of the 44-kDa IvaP intermediate to the fully cleaved 38-kDa form. A trypsin-like serine protease can cleave the inactive form of IvaP. IvaP exhibits strain-specific processing, comparisons of strains C6706, Haiti, E7946, and N1696 Vibrio cholerae serotype O1

Purification (Commentary)

Purification (Comment) Organism
native mature enzyme from cell supernatants by anion exchange chromatography to apparent homogeneity Vibrio cholerae serotype O1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
intelectin + H2O
-
 Vibrio cholerae serotype O1 ?
-
 ?
intelectin + H2O enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae Vibrio cholerae serotype O1 ?
-
 ?
intelectin + H2O
-
 Vibrio cholerae serotype O1 C6706 ?
-
 ?
intelectin + H2O enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae Vibrio cholerae serotype O1 C6706 ?
-
 ?
intelectin + H2O
-
 Vibrio cholerae serotype O1 El Tor Inaba N16961 ?
-
 ?
intelectin + H2O enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae Vibrio cholerae serotype O1 El Tor Inaba N16961 ?
-
 ?
intelectin + H2O
-
 Vibrio cholerae serotype O1 ATCC 39315 ?
-
 ?
intelectin + H2O enzyme IvaP is able to cleave purified intelectin, which inhibits intelectin binding to Vibrio cholerae Vibrio cholerae serotype O1 ATCC 39315 ?
-
 ?
IvaP I9 domain + H2O a temporary inhibitor and substrate of purified IvaP Vibrio cholerae serotype O1 ?
-
 ?
additional information Ser361 contributes to IvaP autoprocessing in stationary-phase cultures Vibrio cholerae serotype O1 ?
-
 ?
additional information enzyme IvaP performs autoprocessing involving residue Ser361. Identification of the N-terminal enzyme IvaP cleavage sequence AAQDNV Vibrio cholerae serotype O1 ?
-
 ?
additional information Ser361 contributes to IvaP autoprocessing in stationary-phase cultures Vibrio cholerae serotype O1 C6706 ?
-
 ?
additional information enzyme IvaP performs autoprocessing involving residue Ser361. Identification of the N-terminal enzyme IvaP cleavage sequence AAQDNV Vibrio cholerae serotype O1 C6706 ?
-
 ?
additional information Ser361 contributes to IvaP autoprocessing in stationary-phase cultures Vibrio cholerae serotype O1 El Tor Inaba N16961 ?
-
 ?
additional information enzyme IvaP performs autoprocessing involving residue Ser361. Identification of the N-terminal enzyme IvaP cleavage sequence AAQDNV Vibrio cholerae serotype O1 El Tor Inaba N16961 ?
-
 ?
additional information Ser361 contributes to IvaP autoprocessing in stationary-phase cultures Vibrio cholerae serotype O1 ATCC 39315 ?
-
 ?
additional information enzyme IvaP performs autoprocessing involving residue Ser361. Identification of the N-terminal enzyme IvaP cleavage sequence AAQDNV Vibrio cholerae serotype O1 ATCC 39315 ?
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Vibrio cholerae serotype O1 N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Vibrio cholerae serotype O1 C6706 N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Vibrio cholerae serotype O1 El Tor Inaba N16961 N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Vibrio cholerae serotype O1 ATCC 39315 N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?

Subunits

Subunits Comment Organism
? x * 49000, recombinant His6-/FLAG-tagged enzyme, SDS-PAGE, x * 47000, recombinant FLAG-tagged enzyme, SDS-PAGE, x * 44000, recombinant detagged enzyme, SDS-PAGE, x * 38000, recombinant detagged, cleaved mature enzyme, SDS-PAGE Vibrio cholerae serotype O1

Synonyms

Synonyms Comment Organism
alkaline serine protease UniProt Vibrio cholerae serotype O1
IvaP
-
 Vibrio cholerae serotype O1
subtilisin-like serine protease
-
 Vibrio cholerae serotype O1
V. cholerae-secreted serine protease
-
 Vibrio cholerae serotype O1
VC_0157
-
 Vibrio cholerae serotype O1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Vibrio cholerae serotype O1

pH Range

pH Minimum pH Maximum Comment Organism
7.5 10 high activity at Vibrio cholerae serotype O1

General Information

General Information Comment Organism
evolution IvaP bears homology to subtilisin-like enzymes, a large family of serine proteases primarily comprised of secreted endopeptidases. IvaP contains a C-terminal bacterial prepeptidase PPC domain that is not typically found in subtilases but facilitates the secretion of other prokaryotic enzymes and is often cleaved extracellularly Vibrio cholerae serotype O1
physiological function Vibrio cholerae-secreted serine protease, IvaP, is active in Vibrio cholerae-infected rabbits and human choleric stool. Enzyme IvaP alters the activity of several host and pathogen enzymes in the gut and, along with other secreted Vibrio cholerae proteases, decreases binding of intelectin, an intestinal carbohydrate-binding protein, to Vibrio cholerae in vivo. IvaP plays a role in modulating intelectin-Vibrio cholerae interactions, it subverts this host-pathogen interaction in vivo Vibrio cholerae serotype O1