2fpq: Difference between revisions

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-[[Image:2fpq.gif|left|200px]]<br /><applet load="2fpq" size="450" color="white" frame="true" align="right" spinBox="true"
-caption="2fpq, resolution 1.65&Aring;" />
-'''Crystal Structure of Botulinum Neurotoxin Type D Light Chain'''<br />
-==Overview==
+==Crystal Structure of Botulinum Neurotoxin Type D Light Chain==
-The seven serotypes (A-G) of botulinum neurotoxins (BoNTs) function, through their proteolytic cleavage of one of three proteins (SNAP-25, Syntaxin, and VAMP) that form the SNARE complex required for synaptic, vesicle fusion. The different BoNTs have very specific protease, recognition requirements, between 15 and 50 amino acids in length, depending on the serotype. However, the structural details involved in, substrate recognition remain largely unknown. Here is reported the 1.65 A, resolution crystal structure of the catalytic domain of BoNT serotype D, (BoNT/D-LC), providing insight into the protein-protein binding, interaction and final proteolysis of VAMP-2. Structural analysis has, identified a hydrophobic pocket potentially involved in substrate, recognition of the P1' VAMP residue (Leu 60) and a second remote site for, recognition of the V1 SNARE motif that is critical for activity. A, structural comparison of BoNT/D-LC with BoNT/F-LC that also recognizes, VAMP-2 one residue away from the BoNT/D-LC site provides additional, molecular details about the unique serotype specific activities. In, particular, BoNT/D prefers a hydrophobic interaction for the V1 motif of, VAMP-2, while BoNT/F adopts a more hydrophilic strategy for recognition of, the same V1 motif.
+<StructureSection load='2fpq' size='340' side='right'caption='[[2fpq]], [[Resolution|resolution]] 1.65&Aring;' scene=''>
+== Structural highlights ==
+<table><tr><td colspan='2'>[[2fpq]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Clostridium_botulinum Clostridium botulinum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2FPQ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2FPQ FirstGlance]. <br>
+</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.65&#8491;</td></tr>
+<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=2fpq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2fpq OCA], [https://pdbe.org/2fpq PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2fpq RCSB], [https://www.ebi.ac.uk/pdbsum/2fpq PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2fpq ProSAT]</span></td></tr>
+</table>
+== Function ==
+[https://www.uniprot.org/uniprot/BXD_CBDP BXD_CBDP] Botulinum toxin causes flaccid paralysis by inhibiting neurotransmitter (acetylcholine) release from the presynaptic membranes of nerve terminals of the eukaryotic host skeletal and autonomic nervous system, with frequent heart or respiratory failure (PubMed:8175689, PubMed:16252491). Precursor of botulinum neurotoxin D for which a proteinaceous coreceptor is controversial. In double SV2A/SV2B knockout mice this toxin does not degrade its synaptobrevin target; introducing SV2A, SV2B or SV2C restores target cleavage (PubMed:21483489). Recognition of SV2 by this toxin does not occur via SV2 glycosylation or its large extracellular loop 4 (PubMed:21483489). Another group does not find a convincing interaction with SV2 (PubMed:21632541). Thus a protein receptor for this BoNT serotype has yet to be definitively proven. Recognizes at least 1 complex polysialylated ganglioside found on neural tissue. Electrical stimulation increases uptake of toxin in an ex vivo assay, presumably by transiently exposing a receptor usually found in eukaryotic target synaptic vesicles (PubMed:19650874, PubMed:21483489, PubMed:21632541). Upon synaptic vesicle recycling the toxin is taken up via the endocytic pathway; when the pH of the toxin-containing endosome drops a structural rearrangement occurs so that the N-terminus of the heavy chain (HC) forms pores that allows the light chain (LC) to translocate into the cytosol (By similarity). Once in the cytosol the disulfide bond linking the 2 subunits is reduced and LC cleaves its target protein on synaptic vesicles, preventing their fusion with the cytoplasmic membrane and thus neurotransmitter release (By similarity). Requires complex eukaryotic host polysialogangliosides for full neurotoxicity and for binding to neurons (PubMed:20704566, PubMed:21483489).[UniProtKB:P0DPI0]<ref>PMID:16252491</ref> <ref>PMID:19650874</ref> <ref>PMID:20704566</ref> <ref>PMID:21483489</ref> <ref>PMID:21632541</ref> <ref>PMID:8175689</ref>   Has proteolytic activity (PubMed:8175689, PubMed:8197120). After translocation into the eukaryotic host cytosol, inhibits neurotransmitter release by acting as a zinc endopeptidase that cleaves the '61-Lys-|-Leu-62' bond of synaptobrevin-1 (VAMP1), and the equivalent 'Lys-|-Leu' sites in VAMP2 and VAMP3 (PubMed:8175689). Cleaves the '49-Lys-|-Ile-50' bond of A.californica synaptobrevin (AC P35589) (PubMed:8197120). This chain probably has to be partially unfolded to translocate into the eukaryotic host cell cytosol (PubMed:15584922).<ref>PMID:8175689</ref> <ref>PMID:8197120</ref> <ref>PMID:15584922</ref>   Responsible for host epithelial cell transcytosis, host nerve cell targeting and translocation of light chain (LC) into eukaryotic host cell cytosol. Composed of 3 subdomains; the translocation domain (TD), and N-terminus and C-terminus of the receptor-binding domain (RBD). The RBD is responsible for the adherence of the toxin to the eukaryotic target cell surface. The N-terminus of the TD wraps an extended belt around the perimeter of the LC, protecting Zn(2+) in the active site; it may also prevent premature LC dissociation from the translocation channel and protect toxin prior to translocation (PubMed:17907800). The TD inserts into synaptic vesicle membrane to allow translocation into the host cytosol (By similarity). The RBD binds eukaryotic host phosphatidylethanolamine, which may serve as toxin receptor (PubMed:16115873). Treatment of synaptosomes with proteinase K does not reduce HC binding, suggesting there is no protein receptor or it is protected from extracellular proteases (PubMed:16115873). HC significantly decreases uptake and toxicity of whole BoNT/D (PubMed:19650874, PubMed:21483489). HC also interferes with uptake of tetanus toxin (PubMed:19650874). Has 2 closely located carbohydrate-binding receptor sites and binds at least 1 GT1b ganglioside (PubMed:20704566). Bind gangliosides in the order GD2 > GT1b > GD1b (PubMed:21632541). Interacts with eukaryotic target protein SV2B (synaptic vesicle glycoprotein 2B) (PubMed:21483489). Expression of SV2A, SV2B or SV2C in mice knocked-out for the SV2 proteins restores entry of BoNT/D and cleavage of VAMP2, suggesting SV2 acts as its receptor (PubMed:21483489). Unlike BoNT/A and BoNT/E, toxin uptake is not mediated by large extracellular loop 4 of SV2 (PubMed:21483489). Another group finds very poor interaction with SV2 proteins, suggesting the possible protein receptor may not have been identified (PubMed:21632541).[UniProtKB:P0DPI0]<ref>PMID:16115873</ref> <ref>PMID:19650874</ref> <ref>PMID:20704566</ref> <ref>PMID:21483489</ref> <ref>PMID:21632541</ref> <ref>PMID:17907800</ref>
+== Evolutionary Conservation ==
+[[Image:Consurf_key_small.gif|200px|right]]
+Check<jmol>
+  <jmolCheckbox>
+    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/fp/2fpq_consurf.spt"</scriptWhenChecked>
+    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
+    <text>to colour the structure by Evolutionary Conservation</text>
+  </jmolCheckbox>
+</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=2fpq ConSurf].
+<div style="clear:both"></div>
-==About this Structure==
+==See Also==
-FPQ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Clostridium_botulinum Clostridium botulinum] with ZN and K as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Bontoxilysin Bontoxilysin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.24.69 3.4.24.69] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2FPQ OCA].
+*[[Botulinum neurotoxin 3D structures|Botulinum neurotoxin 3D structures]]
+== References ==
-==Reference==
+<references/>
-Structure of botulinum neurotoxin type D light chain at 1.65 A resolution: repercussions for VAMP-2 substrate specificity., Arndt JW, Chai Q, Christian T, Stevens RC, Biochemistry. 2006 Mar 14;45(10):3255-62. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16519520 16519520]
+__TOC__
-[[Category: Bontoxilysin]]
+</StructureSection>
 [[Category: Clostridium botulinum]]
-[[Category: Single protein]]
+[[Category: Large Structures]]
-[[Category: Arndt, J.W.]]
+[[Category: Arndt JW]]
-[[Category: Chai, Q.]]
+[[Category: Chai Q]]
-[[Category: Christian, T.]]
+[[Category: Christian T]]
-[[Category: Stevens, R.C.]]
+[[Category: Stevens RC]]
-[[Category: K]]
-[[Category: ZN]]
-[[Category: hexxh metalloprotease]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 10:41:20 2007''