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Literature summary for 7.4.2.8 extracted from

  • Yoshida, Y.; Miki, T.; Ono, S.; Haneda, T.; Ito, M.; Okada, N.
    Functional characterization of the type III secretion ATPase SsaN encoded by Salmonella pathogenicity island 2 (2014), PLoS ONE, 9, e94347.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 7.4.2.8

Cloned(Commentary)

Cloned (Comment) Organism
gene ssaN, recombinant expression of wild-type and mutant enzymes tagged with hemagglutinin, c-Myc, a FLAG- or a His6-tag, in Escherichia coli and Salmonella enterica, co-expression with SsaK-2HA and SsaQ-2HA fusion proteins Salmonella enterica

Protein Variants

Protein Variants Comment Organism
R192G site-directed mutagenesis of the conserved arginine residue at position 192 of SsaN located in the dicyclohexylcarbodiimide-binding site in the catalytic domain, inactive mutant showing no ATPase and translocation activity. Introducing a plasmid that expresses SsaNR192G into the DELTAssaN mutant fails to complement SseJ secretion. Mutant SsaNR192G-Myc-His6 binds to chaperone SsaE Salmonella enterica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.81
-
 ATP purified recombinant SsaN-Myc-His6 enzyme, pH 8.0, 37°C Salmonella enterica

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane SsaN and its complex colocalize to the membrane fraction under type III secretion system T3SS-2 inducing conditions, SsaN, SsaK, and SsaQ co-localize to bacterial membranes after T3SS activation Salmonella enterica 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Salmonella enterica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + H2O Salmonella enterica
-
 ADP + phosphate
-
 ?
ATP + H2O Salmonella enterica SL1344
-
 ADP + phosphate
-
 ?
additional information Salmonella enterica enzyme SsaN binds to Salmonella pathogenicity island 2 (SPI-2) specific chaperones, including SsaE, SseA, SscA, and SscB that facilitate translocator/effector secretion, SsaN dissociates a chaperone-effector complex, SsaE and SseB, in an ATP-dependent manner. Effector release is dependent on a conserved arginine residue at position 192 of SsaN, and this is essential for its enzymatic activity ?
-
 ?
additional information Salmonella enterica SL1344 enzyme SsaN binds to Salmonella pathogenicity island 2 (SPI-2) specific chaperones, including SsaE, SseA, SscA, and SscB that facilitate translocator/effector secretion, SsaN dissociates a chaperone-effector complex, SsaE and SseB, in an ATP-dependent manner. Effector release is dependent on a conserved arginine residue at position 192 of SsaN, and this is essential for its enzymatic activity ?
-
 ?

Organism

Organism UniProt Comment Textmining
Salmonella enterica A0A0H3NB84 serovar Typhimurium, gene ssaN
-
Salmonella enterica SL1344 A0A0H3NB84 serovar Typhimurium, gene ssaN
-

Purification (Commentary)

Purification (Comment) Organism
recombinant SsaN-Myc-His6, EscN-Myc-His6, and SsaNR192G-Myc-His6 wefrom Escherichia coli by nickel affinity chromatography Salmonella enterica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.36
-
 purified recombinant SsaN-Myc-His6 enzyme, pH 8.0, 37°C Salmonella enterica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O
-
 Salmonella enterica ADP + phosphate
-
 ?
ATP + H2O malachite green reagent assay for activity determination Salmonella enterica ADP + phosphate
-
 ?
ATP + H2O
-
 Salmonella enterica SL1344 ADP + phosphate
-
 ?
ATP + H2O malachite green reagent assay for activity determination Salmonella enterica SL1344 ADP + phosphate
-
 ?
additional information enzyme SsaN binds to Salmonella pathogenicity island 2 (SPI-2) specific chaperones, including SsaE, SseA, SscA, and SscB that facilitate translocator/effector secretion, SsaN dissociates a chaperone-effector complex, SsaE and SseB, in an ATP-dependent manner. Effector release is dependent on a conserved arginine residue at position 192 of SsaN, and this is essential for its enzymatic activity Salmonella enterica ?
-
 ?
additional information enzyme SsaN binds to Salmonella pathogenicity island 2 (SPI-2) specific chaperones, including SsaE, SseA, SscA, and SscB that facilitate translocator/effector secretion, SsaN dissociates a chaperone-effector complex, SsaE and SseB, in an ATP-dependent manner. Effector release is dependent on a conserved arginine residue at position 192 of SsaN, and this is essential for its enzymatic activity Salmonella enterica SL1344 ?
-
 ?

Synonyms

Synonyms Comment Organism
SsaN
-
 Salmonella enterica
type III secretion ATPase
-
 Salmonella enterica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Salmonella enterica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Salmonella enterica

General Information

General Information Comment Organism
metabolism type III secretion system T3SS-1 facilitates host cell invasion and inflammation, whereas type III secretion system T3SS-2 mediates intracellular survival and immune evasion Salmonella enterica
additional information the conserved arginine residue at position 192 of SsaN located in the dicyclohexylcarbodiimide-binding site (DCCD box) in the catalytic domain is essential for ATPase activity Salmonella enterica
physiological function enzyme SsaN hydrolyzes ATP in vitro and is essential for type III secretion system, T3SS, function and Salmonella virulence in vivo. Protein-protein interaction analyses reveal that SsaN interacts with SsaK and SsaQ to form the C ring complex. The T3SS-2-associated ATPase SsaN contributes to T3SS-2 effector translocation efficiency. Enzyme SsaN is required for the export of another T3SS-2 effector, SseJ, which is encoded outside of the T3SS-2 region. Enzyme SsaN releases the translocator protein SseB from the T3SS-2 specific chaperone SsaE in an ATP-dependent manner. Gene ssaN contributes to Salmonella virulence in the mouse model of systemic infection Salmonella enterica