EC Number | Cloned (Comment) | Organism |
---|---|---|
7.4.2.8 | 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 |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
7.4.2.8 | 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 |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.4.2.8 | 0.81 | - |
ATP | purified recombinant SsaN-Myc-His6 enzyme, pH 8.0, 37°C | Salmonella enterica |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
7.4.2.8 | 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 | - |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
7.4.2.8 | Mg2+ | required | Salmonella enterica |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.4.2.8 | ATP + H2O | Salmonella enterica | - |
ADP + phosphate | - |
? | |
7.4.2.8 | ATP + H2O | Salmonella enterica SL1344 | - |
ADP + phosphate | - |
? | |
7.4.2.8 | 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 | ? | - |
? | |
7.4.2.8 | 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 | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
7.4.2.8 | Salmonella enterica | A0A0H3NB84 | serovar Typhimurium, gene ssaN | - |
7.4.2.8 | Salmonella enterica SL1344 | A0A0H3NB84 | serovar Typhimurium, gene ssaN | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
7.4.2.8 | recombinant SsaN-Myc-His6, EscN-Myc-His6, and SsaNR192G-Myc-His6 wefrom Escherichia coli by nickel affinity chromatography | Salmonella enterica |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
7.4.2.8 | 0.36 | - |
purified recombinant SsaN-Myc-His6 enzyme, pH 8.0, 37°C | Salmonella enterica |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.4.2.8 | ATP + H2O | - |
Salmonella enterica | ADP + phosphate | - |
? | |
7.4.2.8 | ATP + H2O | malachite green reagent assay for activity determination | Salmonella enterica | ADP + phosphate | - |
? | |
7.4.2.8 | ATP + H2O | - |
Salmonella enterica SL1344 | ADP + phosphate | - |
? | |
7.4.2.8 | ATP + H2O | malachite green reagent assay for activity determination | Salmonella enterica SL1344 | ADP + phosphate | - |
? | |
7.4.2.8 | 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 | ? | - |
? | |
7.4.2.8 | 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 | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
7.4.2.8 | SsaN | - |
Salmonella enterica |
7.4.2.8 | type III secretion ATPase | - |
Salmonella enterica |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
7.4.2.8 | 37 | - |
assay at | Salmonella enterica |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
7.4.2.8 | 8 | - |
assay at | Salmonella enterica |
EC Number | General Information | Comment | Organism |
---|---|---|---|
7.4.2.8 | 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 |
7.4.2.8 | 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 |
7.4.2.8 | 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 |