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| | ==Crystal Structure of ATPase delta1-79 Spa47 K165A== | | ==Crystal Structure of ATPase delta1-79 Spa47 K165A== |
| - | <StructureSection load='5syp' size='340' side='right' caption='[[5syp]], [[Resolution|resolution]] 2.15Å' scene=''> | + | <StructureSection load='5syp' size='340' side='right'caption='[[5syp]], [[Resolution|resolution]] 2.15Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5syp]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"shigella_paradysenteriae"_weldin_1927 "shigella paradysenteriae" weldin 1927]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5SYP OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5SYP FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5syp]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Shigella_flexneri Shigella flexneri]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5SYP OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5SYP FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.15Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5swj|5swj]], [[5swl|5swl]], [[5syr|5syr]]</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">spaL, mxiB, spa47, CP0149 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=623 "Shigella paradysenteriae" Weldin 1927])</td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5syp FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5syp OCA], [https://pdbe.org/5syp PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5syp RCSB], [https://www.ebi.ac.uk/pdbsum/5syp PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5syp ProSAT]</span></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/H(+)-transporting_two-sector_ATPase H(+)-transporting two-sector ATPase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.6.3.14 3.6.3.14] </span></td></tr>
| + | |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5syp FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5syp OCA], [http://pdbe.org/5syp PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5syp RCSB], [http://www.ebi.ac.uk/pdbsum/5syp PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5syp ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/SPAL_SHIFL SPAL_SHIFL]] Required for surface presentation of invasion plasmid antigens. Probable catalytic subunit of a protein translocase. Required for invasion and for secretion of the three IPA proteins. | + | [https://www.uniprot.org/uniprot/SCTN_SHIFL SCTN_SHIFL] ATPase component of the type III secretion system (T3SS), also called injectisome, which is used to inject bacterial effector proteins into eukaryotic host cells (PubMed:26947936, PubMed:27770024, PubMed:29595954). Acts as a molecular motor to provide the energy that is required for the export of proteins (Probable). Required for type III secretion apparatus (T3SA) formation, proper protein secretion, host cell invasion and virulence (PubMed:26947936, PubMed:27770024, PubMed:31162724). May play a critical role in T3SS substrate recognition, disassembly of the effector/chaperone complex and unfolding of the effector in an ATP-dependent manner prior to secretion (By similarity).[UniProtKB:P0A1B9]<ref>PMID:26947936</ref> <ref>PMID:27770024</ref> <ref>PMID:29595954</ref> <ref>PMID:31162724</ref> <ref>PMID:26947936</ref> <ref>PMID:27770024</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Shigella paradysenteriae weldin 1927]] | + | [[Category: Large Structures]] |
| - | [[Category: Burgess, J L]] | + | [[Category: Shigella flexneri]] |
| - | [[Category: Burgess, R A]] | + | [[Category: Burgess JL]] |
| - | [[Category: Dickenson, N E]] | + | [[Category: Burgess RA]] |
| - | [[Category: Johnson, S J]] | + | [[Category: Dickenson NE]] |
| - | [[Category: Morales, Y]] | + | [[Category: Johnson SJ]] |
| - | [[Category: Atpase]] | + | [[Category: Morales Y]] |
| - | [[Category: Hydrolase]]
| + | |
| Structural highlights
Function
SCTN_SHIFL ATPase component of the type III secretion system (T3SS), also called injectisome, which is used to inject bacterial effector proteins into eukaryotic host cells (PubMed:26947936, PubMed:27770024, PubMed:29595954). Acts as a molecular motor to provide the energy that is required for the export of proteins (Probable). Required for type III secretion apparatus (T3SA) formation, proper protein secretion, host cell invasion and virulence (PubMed:26947936, PubMed:27770024, PubMed:31162724). May play a critical role in T3SS substrate recognition, disassembly of the effector/chaperone complex and unfolding of the effector in an ATP-dependent manner prior to secretion (By similarity).[UniProtKB:P0A1B9][1] [2] [3] [4] [5] [6]
Publication Abstract from PubMed
Like many Gram-negative pathogens, Shigella rely on a complex type three secretion system (T3SS) to inject effector proteins into host cells, take over host functions, and ultimately establish infection. Despite these critical roles, the energetics and regulatory mechanisms controlling the T3SS and pathogen virulence remain largely unclear. In this study, we present a series of high-resolution crystal structures of Spa47 and use the structures to model an activated Spa47 oligomer, finding that ATP hydrolysis may be supported by specific sidechain contributions from adjacent protomers within the complex. Follow-up mutagenesis experiments targeting the predicted active site residues validate the oligomeric model and determined that each of the tested residues are essential for Spa47 ATPase activity, though they are not directly responsible for stable oligomer formation. While an N-terminal domain truncation was necessary for crystal formation, it resulted in strictly monomeric Spa47 that is unable to hydrolyze ATP, despite maintaining the canonical ATPase core structure and active site residues. Coupled with studies of ATPase inactive full-length Spa47 point mutants, we find that Spa47 oligomerization and ATP hydrolysis are needed for complete T3SS apparatus formation, a proper translocator secretion profile, and Shigella virulence. This work represents the first structure-function characterization of Spa47, uniquely complementing the multitude of included Shigella T3SS phenotype assays and providing a more complete understanding of T3SS ATPase-mediated pathogen virulence. Additionally, these findings provide a strong platform for follow-up studies evaluating regulation of Spa47 oligomerization in vivo as a much needed means of treating and perhaps preventing shigellosis.
Structural and Biochemical Characterization of Spa47 Provides Mechanistic Insight into Type III Secretion System ATPase Activation and Shigella Virulence Regulation.,Burgess JL, Burgess RA, Morales Y, Bouvang JM, Johnson SJ, Dickenson NE J Biol Chem. 2016 Oct 21. pii: jbc.M116.755256. PMID:27770024[7]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Burgess JL, Jones HB, Kumar P, Toth RT 4th, Middaugh CR, Antony E, Dickenson NE. Spa47 is an oligomerization-activated type three secretion system (T3SS) ATPase from Shigella flexneri. Protein Sci. 2016 May;25(5):1037-48. PMID:26947936 doi:10.1002/pro.2917
- ↑ Burgess JL, Burgess RA, Morales Y, Bouvang JM, Johnson SJ, Dickenson NE. Structural and Biochemical Characterization of Spa47 Provides Mechanistic Insight into Type III Secretion System ATPase Activation and Shigella Virulence Regulation. J Biol Chem. 2016 Oct 21. pii: jbc.M116.755256. PMID:27770024 doi:http://dx.doi.org/10.1074/jbc.M116.755256
- ↑ Case HB, Dickenson NE. MxiN Differentially Regulates Monomeric and Oligomeric Species of the Shigella Type Three Secretion System ATPase Spa47. Biochemistry. 2018 Apr 17;57(15):2266-2277. PMID:29595954 doi:10.1021/acs.biochem.8b00070
- ↑ Demler HJ, Case HB, Morales Y, Bernard AR, Johnson SJ, Dickenson NE. Interfacial amino acids support Spa47 oligomerization and shigella type three secretion system activation. Proteins. 2019 Jun 4. doi: 10.1002/prot.25754. PMID:31162724 doi:http://dx.doi.org/10.1002/prot.25754
- ↑ Burgess JL, Jones HB, Kumar P, Toth RT 4th, Middaugh CR, Antony E, Dickenson NE. Spa47 is an oligomerization-activated type three secretion system (T3SS) ATPase from Shigella flexneri. Protein Sci. 2016 May;25(5):1037-48. PMID:26947936 doi:10.1002/pro.2917
- ↑ Burgess JL, Burgess RA, Morales Y, Bouvang JM, Johnson SJ, Dickenson NE. Structural and Biochemical Characterization of Spa47 Provides Mechanistic Insight into Type III Secretion System ATPase Activation and Shigella Virulence Regulation. J Biol Chem. 2016 Oct 21. pii: jbc.M116.755256. PMID:27770024 doi:http://dx.doi.org/10.1074/jbc.M116.755256
- ↑ Burgess JL, Burgess RA, Morales Y, Bouvang JM, Johnson SJ, Dickenson NE. Structural and Biochemical Characterization of Spa47 Provides Mechanistic Insight into Type III Secretion System ATPase Activation and Shigella Virulence Regulation. J Biol Chem. 2016 Oct 21. pii: jbc.M116.755256. PMID:27770024 doi:http://dx.doi.org/10.1074/jbc.M116.755256
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