2iy9

From Proteopedia

(Difference between revisions)

Revision as of 14:29, 13 December 2023

Crystal structure of the A-subunit of the AB5 toxin from E. coli

Structural highlights

2iy9 is a 1 chain structure with sequence from Escherichia coli. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 1.8Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

SUBA_ECOLX Protease subunit of subtilase cytotoxin SubAB5 (PubMed:17024087). An endoprotease specific for host endoplasmic reticulum (ER) chaperone BiP/HSPA5, has no activity on human HSP70 or HSPA8 (PubMed:17024087). Cleaves between 'Leu-416' and 'Leu-417' of BiP/HSPA5 in the hinge between BiP's ATPase and protein-binding domains (PubMed:17024087). This induces host ER stress response and eventual cell death (PubMed:18005237, PubMed:18433465). Culture supernatant of E.coli expressing both subA and subB are toxic for Vero cells (African green monkey kidney cell line), Chinese hamster ovary cells and Hct-8 cells (human colonic epithelial cell line); the subunits are not toxic individually (PubMed:15226357). Purified SubAB5 is highly toxic, <0.1 pg is able to kill at least 50% of 30'000 Vero cells in a microtiter plate assay after 3 days; no cytotoxicity is seen at 24 hours (PubMed:15226357). Preabsorption with cells expressing a ganglioside GM2 mimic reduced cytotoxicity of SubAB5 by 93% in the Vero cytotoxicity assay (PubMed:15226357). Intraperitoneal injection of 200 ng of purified SubAB5 kills mice; the higher the dose the faster the mice die. Animals injected intraperitoneally with purified SubAB5 have microvascular thrombi in the brain and other organs, including the renal tubules and glomeruli (PubMed:15226357). Injection induces an unfolded response in mice (PubMed:17024087). Mice fed E.coli cells expressing cloned SubAB5 experience drastic weight loss and appear ill and lethargic (PubMed:15226357). Protein synthesis in Vero cells is transiently inhibited by SubAB5; both subunits are required for this effect (PubMed:17101670, PubMed:18005237, PubMed:18433465). Inhibition of protein synthesis is prevented by brefeldin A; cells are arrested in the G1 phase (PubMed:18005237). SubAB5 at 100 ng/ml induced caspase-dependent apoptosis in Vero cells through mitochondrial membrane damage (PubMed:19380466).^[1] ^[2] ^[3] ^[4] ^[5] ^[6]

Evolutionary Conservation

Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.

Publication Abstract from PubMed

AB5 toxins are produced by pathogenic bacteria and consist of enzymatic A subunits that corrupt essential eukaryotic cell functions, and pentameric B subunits that mediate uptake into the target cell. AB5 toxins include the Shiga, cholera and pertussis toxins and a recently discovered fourth family, subtilase cytotoxin, which is produced by certain Shiga toxigenic strains of Escherichia coli. Here we show that the extreme cytotoxicity of this toxin for eukaryotic cells is due to a specific single-site cleavage of the essential endoplasmic reticulum chaperone BiP/GRP78. The A subunit is a subtilase-like serine protease; structural studies revealed an unusually deep active-site cleft, which accounts for its exquisite substrate specificity. A single amino-acid substitution in the BiP target site prevented cleavage, and co-expression of this resistant protein protected transfected cells against the toxin. BiP is a master regulator of endoplasmic reticulum function, and its cleavage by subtilase cytotoxin represents a previously unknown trigger for cell death.

AB5 subtilase cytotoxin inactivates the endoplasmic reticulum chaperone BiP.,Paton AW, Beddoe T, Thorpe CM, Whisstock JC, Wilce MC, Rossjohn J, Talbot UM, Paton JC Nature. 2006 Oct 5;443(7111):548-52. PMID:17024087^[7]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Paton AW, Srimanote P, Talbot UM, Wang H, Paton JC. A new family of potent AB(5) cytotoxins produced by Shiga toxigenic Escherichia coli. J Exp Med. 2004 Jul 5;200(1):35-46. doi: 10.1084/jem.20040392. Epub 2004 Jun 28. PMID:15226357 doi:http://dx.doi.org/10.1084/jem.20040392
↑ Paton AW, Beddoe T, Thorpe CM, Whisstock JC, Wilce MC, Rossjohn J, Talbot UM, Paton JC. AB5 subtilase cytotoxin inactivates the endoplasmic reticulum chaperone BiP. Nature. 2006 Oct 5;443(7111):548-52. PMID:17024087 doi:10.1038/nature05124
↑ Morinaga N, Yahiro K, Matsuura G, Watanabe M, Nomura F, Moss J, Noda M. Two distinct cytotoxic activities of subtilase cytotoxin produced by shiga-toxigenic Escherichia coli. Infect Immun. 2007 Jan;75(1):488-96. doi: 10.1128/IAI.01336-06. Epub 2006 Nov 13. PMID:17101670 doi:http://dx.doi.org/10.1128/IAI.01336-06
↑ Morinaga N, Yahiro K, Matsuura G, Moss J, Noda M. Subtilase cytotoxin, produced by Shiga-toxigenic Escherichia coli, transiently inhibits protein synthesis of Vero cells via degradation of BiP and induces cell cycle arrest at G1 by downregulation of cyclin D1. Cell Microbiol. 2008 Apr;10(4):921-9. doi: 10.1111/j.1462-5822.2007.01094.x. Epub, 2007 Nov 14. PMID:18005237 doi:http://dx.doi.org/10.1111/j.1462-5822.2007.01094.x
↑ Wolfson JJ, May KL, Thorpe CM, Jandhyala DM, Paton JC, Paton AW. Subtilase cytotoxin activates PERK, IRE1 and ATF6 endoplasmic reticulum stress-signalling pathways. Cell Microbiol. 2008 Sep;10(9):1775-86. doi: 10.1111/j.1462-5822.2008.01164.x., Epub 2008 Apr 21. PMID:18433465 doi:http://dx.doi.org/10.1111/j.1462-5822.2008.01164.x
↑ Matsuura G, Morinaga N, Yahiro K, Komine R, Moss J, Yoshida H, Noda M. Novel subtilase cytotoxin produced by Shiga-toxigenic Escherichia coli induces apoptosis in vero cells via mitochondrial membrane damage. Infect Immun. 2009 Jul;77(7):2919-24. doi: 10.1128/IAI.01510-08. Epub 2009 Apr, 20. PMID:19380466 doi:http://dx.doi.org/10.1128/IAI.01510-08
↑ Paton AW, Beddoe T, Thorpe CM, Whisstock JC, Wilce MC, Rossjohn J, Talbot UM, Paton JC. AB5 subtilase cytotoxin inactivates the endoplasmic reticulum chaperone BiP. Nature. 2006 Oct 5;443(7111):548-52. PMID:17024087 doi:10.1038/nature05124

1 Structural highlights
2 Function
3 Evolutionary Conservation
4 Publication Abstract from PubMed
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/2iy9"

@@ Line 3: / Line 3: @@
 <StructureSection load='2iy9' size='340' side='right'caption='[[2iy9]], [[Resolution|resolution]] 1.80&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>[[2iy9]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/"bacillus_coli"_migula_1895 "bacillus coli" migula 1895]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2IY9 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2IY9 FirstGlance]. <br>
+<table><tr><td colspan='2'>[[2iy9]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2IY9 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2IY9 FirstGlance]. <br>
-</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=2iy9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2iy9 OCA], [https://pdbe.org/2iy9 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2iy9 RCSB], [https://www.ebi.ac.uk/pdbsum/2iy9 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2iy9 ProSAT]</span></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]] 1.8&#8491;</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=2iy9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2iy9 OCA], [https://pdbe.org/2iy9 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2iy9 RCSB], [https://www.ebi.ac.uk/pdbsum/2iy9 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2iy9 ProSAT]</span></td></tr>
 </table>
+== Function ==
+[https://www.uniprot.org/uniprot/SUBA_ECOLX SUBA_ECOLX] Protease subunit of subtilase cytotoxin SubAB5 (PubMed:17024087). An endoprotease specific for host endoplasmic reticulum (ER) chaperone BiP/HSPA5, has no activity on human HSP70 or HSPA8 (PubMed:17024087). Cleaves between 'Leu-416' and 'Leu-417' of BiP/HSPA5 in the hinge between BiP's ATPase and protein-binding domains (PubMed:17024087). This induces host ER stress response and eventual cell death (PubMed:18005237, PubMed:18433465). Culture supernatant of E.coli expressing both subA and subB are toxic for Vero cells (African green monkey kidney cell line), Chinese hamster ovary cells and Hct-8 cells (human colonic epithelial cell line); the subunits are not toxic individually (PubMed:15226357). Purified SubAB5 is highly toxic, <0.1 pg is able to kill at least 50% of 30'000 Vero cells in a microtiter plate assay after 3 days; no cytotoxicity is seen at 24 hours (PubMed:15226357). Preabsorption with cells expressing a ganglioside GM2 mimic reduced cytotoxicity of SubAB5 by 93% in the Vero cytotoxicity assay (PubMed:15226357). Intraperitoneal injection of 200 ng of purified SubAB5 kills mice; the higher the dose the faster the mice die. Animals injected intraperitoneally with purified SubAB5 have microvascular thrombi in the brain and other organs, including the renal tubules and glomeruli (PubMed:15226357). Injection induces an unfolded response in mice (PubMed:17024087). Mice fed E.coli cells expressing cloned SubAB5 experience drastic weight loss and appear ill and lethargic (PubMed:15226357). Protein synthesis in Vero cells is transiently inhibited by SubAB5; both subunits are required for this effect (PubMed:17101670, PubMed:18005237, PubMed:18433465). Inhibition of protein synthesis is prevented by brefeldin A; cells are arrested in the G1 phase (PubMed:18005237). SubAB5 at 100 ng/ml induced caspase-dependent apoptosis in Vero cells through mitochondrial membrane damage (PubMed:19380466).<ref>PMID:15226357</ref> <ref>PMID:17024087</ref> <ref>PMID:17101670</ref> <ref>PMID:18005237</ref> <ref>PMID:18433465</ref> <ref>PMID:19380466</ref>
 == Evolutionary Conservation ==
 [[Image:Consurf_key_small.gif|200px|right]]
@@ Line 29: / Line 32: @@
 __TOC__
 </StructureSection>
-[[Category: Bacillus coli migula 1895]]
+[[Category: Escherichia coli]]
 [[Category: Large Structures]]
-[[Category: Beddoe, T]]
+[[Category: Beddoe T]]
-[[Category: Paton, A W]]
+[[Category: Paton AW]]
-[[Category: Paton, J C]]
+[[Category: Paton JC]]
-[[Category: Rossjohn, J]]
+[[Category: Rossjohn J]]
-[[Category: Talbot, U M]]
+[[Category: Talbot UM]]
-[[Category: Thorpe, C M]]
+[[Category: Thorpe CM]]
-[[Category: Whisstock, J C]]
+[[Category: Whisstock JC]]
-[[Category: Wilce, M C.J]]
+[[Category: Wilce MCJ]]
-[[Category: Coli]]
-[[Category: Plasmid]]
-[[Category: Shiga]]
-[[Category: Toxin]]

2iy9

From Proteopedia

Revision as of 14:29, 13 December 2023

Crystal structure of the A-subunit of the AB5 toxin from E. coli

Structural highlights

Function

Evolutionary Conservation

Publication Abstract from PubMed

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

Views

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