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| - | [[Image:1war.gif|left|200px]]<br /> | |
| - | <applet load="1war" size="450" color="white" frame="true" align="right" spinBox="true" | |
| - | caption="1war, resolution 2.22Å" /> | |
| - | '''RECOMBINANT HUMAN PURPLE ACID PHOSPHATASE EXPRESSED IN PICHIA PASTORIS'''<br /> | |
| | | | |
| - | ==Overview== | + | ==Recombinant Human Purple Acid Phosphatase expressed in Pichia Pastoris== |
| - | The crystal structure of human purple acid phosphatase recombinantly, expressed in Escherichia coli (rHPAP(Ec)) and Pichia pastoris (rHPAP(Pp)), has been determined in two different crystal forms, both at 2.2A, resolution. In both cases, the enzyme crystallized in its oxidized, (inactive) state, in which both Fe atoms in the dinuclear active site are, Fe(III). The main difference between the two structures is the, conformation of the enzyme "repression loop". Proteolytic cleavage of this, loop in vivo or in vitro results in significant activation of the, mammalian PAPs. In the crystals obtained from rHPAP(Ec), the carboxylate, side-chain of Asp145 of this loop acts as a bidentate ligand that bridges, the two metal atoms, in a manner analogous to a possible binding mode for, a phosphate ester substrate in the enzyme-substrate complex. The, carboxylate side-chain of Asp145 and the neighboring Phe146 side-chain, thus block the active site, thereby inactivating the enzyme. In the, crystal structure of rHPAP(Pp), the enzyme "repression loop" has an open, conformation similar to that observed in other mammalian PAP structures., The present structures demonstrate that the repression loop exhibits, significant conformational flexibility, and the observed alternate binding, mode suggests a possible inhibitory role for this loop. | + | <StructureSection load='1war' size='340' side='right'caption='[[1war]], [[Resolution|resolution]] 2.22Å' scene=''> |
| | + | == Structural highlights == |
| | + | <table><tr><td colspan='2'>[[1war]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1WAR OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1WAR 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]] 2.22Å</td></tr> |
| | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FE:FE+(III)+ION'>FE</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</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=1war FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1war OCA], [https://pdbe.org/1war PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1war RCSB], [https://www.ebi.ac.uk/pdbsum/1war PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1war ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Disease == |
| | + | [https://www.uniprot.org/uniprot/PPA5_HUMAN PPA5_HUMAN] Defects in ACP5 are the cause of spondyloenchondrodysplasia with immune dysregulation (SPENCDI) [MIM:[https://omim.org/entry/607944 607944]. A disease characterized by vertebral and metaphyseal dysplasia, spasticity with cerebral calcifications, and strong predisposition to autoimmune diseases. The skeletal dysplasia is characterized by radiolucent and irregular spondylar and metaphyseal lesions that represent islands of chondroid tissue within bone. Note=ACP5 inactivating mutations result in a functional excess of phosphorylated osteopontin causing deregulation of osteopontin signaling and consequential autoimmune disease.<ref>PMID:21217755</ref> <ref>PMID:21217752</ref> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/PPA5_HUMAN PPA5_HUMAN] Involved in osteopontin/bone sialoprotein dephosphorylation. Its expression seems to increase in certain pathological states such as Gaucher and Hodgkin diseases, the hairy cell, the B-cell, and the T-cell leukemias. |
| | + | == 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/wa/1war_consurf.spt"</scriptWhenChecked> |
| | + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.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=1war ConSurf]. |
| | + | <div style="clear:both"></div> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | The crystal structure of human purple acid phosphatase recombinantly expressed in Escherichia coli (rHPAP(Ec)) and Pichia pastoris (rHPAP(Pp)) has been determined in two different crystal forms, both at 2.2A resolution. In both cases, the enzyme crystallized in its oxidized (inactive) state, in which both Fe atoms in the dinuclear active site are Fe(III). The main difference between the two structures is the conformation of the enzyme "repression loop". Proteolytic cleavage of this loop in vivo or in vitro results in significant activation of the mammalian PAPs. In the crystals obtained from rHPAP(Ec), the carboxylate side-chain of Asp145 of this loop acts as a bidentate ligand that bridges the two metal atoms, in a manner analogous to a possible binding mode for a phosphate ester substrate in the enzyme-substrate complex. The carboxylate side-chain of Asp145 and the neighboring Phe146 side-chain thus block the active site, thereby inactivating the enzyme. In the crystal structure of rHPAP(Pp), the enzyme "repression loop" has an open conformation similar to that observed in other mammalian PAP structures. The present structures demonstrate that the repression loop exhibits significant conformational flexibility, and the observed alternate binding mode suggests a possible inhibitory role for this loop. |
| | | | |
| - | ==About this Structure==
| + | Crystal structures of recombinant human purple Acid phosphatase with and without an inhibitory conformation of the repression loop.,Strater N, Jasper B, Scholte M, Krebs B, Duff AP, Langley DB, Han R, Averill BA, Freeman HC, Guss JM J Mol Biol. 2005 Aug 5;351(1):233-46. Epub 2005 Apr 26. PMID:15993892<ref>PMID:15993892</ref> |
| - | 1WAR is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with NAG, FE and PO4 as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Acid_phosphatase Acid phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.2 3.1.3.2] Structure known Active Site: AC1. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1WAR OCA].
| + | |
| | | | |
| - | ==Reference==
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| - | Crystal structures of recombinant human purple Acid phosphatase with and without an inhibitory conformation of the repression loop., Strater N, Jasper B, Scholte M, Krebs B, Duff AP, Langley DB, Han R, Averill BA, Freeman HC, Guss JM, J Mol Biol. 2005 Aug 5;351(1):233-46. Epub 2005 Apr 26. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=15993892 15993892]
| + | </div> |
| - | [[Category: Acid phosphatase]]
| + | <div class="pdbe-citations 1war" style="background-color:#fffaf0;"></div> |
| - | [[Category: Homo sapiens]]
| + | |
| - | [[Category: Single protein]]
| + | |
| - | [[Category: Averill, B.A.]]
| + | |
| - | [[Category: Duff, A.P.]]
| + | |
| - | [[Category: Freeman, H.C.]]
| + | |
| - | [[Category: Guss, J.M.]]
| + | |
| - | [[Category: Han, R.]]
| + | |
| - | [[Category: Langley, D.B.]]
| + | |
| - | [[Category: FE]]
| + | |
| - | [[Category: NAG]]
| + | |
| - | [[Category: PO4]]
| + | |
| - | [[Category: glycoprotein]]
| + | |
| - | [[Category: hydrolase]]
| + | |
| - | [[Category: iron]]
| + | |
| - | [[Category: iron transport]]
| + | |
| - | [[Category: metalloenzyme]]
| + | |
| - | [[Category: purple acid phosphatase]]
| + | |
| - | [[Category: signal]]
| + | |
| - | [[Category: tartrate resistant acid phosphatase]]
| + | |
| - | [[Category: uteroferrin]]
| + | |
| | | | |
| - | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 17:30:43 2007''
| + | ==See Also== |
| | + | *[[Acid phosphatase 3D structures|Acid phosphatase 3D structures]] |
| | + | == References == |
| | + | <references/> |
| | + | __TOC__ |
| | + | </StructureSection> |
| | + | [[Category: Homo sapiens]] |
| | + | [[Category: Large Structures]] |
| | + | [[Category: Averill BA]] |
| | + | [[Category: Duff AP]] |
| | + | [[Category: Freeman HC]] |
| | + | [[Category: Guss JM]] |
| | + | [[Category: Han R]] |
| | + | [[Category: Langley DB]] |
| Structural highlights
Disease
PPA5_HUMAN Defects in ACP5 are the cause of spondyloenchondrodysplasia with immune dysregulation (SPENCDI) [MIM:607944. A disease characterized by vertebral and metaphyseal dysplasia, spasticity with cerebral calcifications, and strong predisposition to autoimmune diseases. The skeletal dysplasia is characterized by radiolucent and irregular spondylar and metaphyseal lesions that represent islands of chondroid tissue within bone. Note=ACP5 inactivating mutations result in a functional excess of phosphorylated osteopontin causing deregulation of osteopontin signaling and consequential autoimmune disease.[1] [2]
Function
PPA5_HUMAN Involved in osteopontin/bone sialoprotein dephosphorylation. Its expression seems to increase in certain pathological states such as Gaucher and Hodgkin diseases, the hairy cell, the B-cell, and the T-cell leukemias.
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
The crystal structure of human purple acid phosphatase recombinantly expressed in Escherichia coli (rHPAP(Ec)) and Pichia pastoris (rHPAP(Pp)) has been determined in two different crystal forms, both at 2.2A resolution. In both cases, the enzyme crystallized in its oxidized (inactive) state, in which both Fe atoms in the dinuclear active site are Fe(III). The main difference between the two structures is the conformation of the enzyme "repression loop". Proteolytic cleavage of this loop in vivo or in vitro results in significant activation of the mammalian PAPs. In the crystals obtained from rHPAP(Ec), the carboxylate side-chain of Asp145 of this loop acts as a bidentate ligand that bridges the two metal atoms, in a manner analogous to a possible binding mode for a phosphate ester substrate in the enzyme-substrate complex. The carboxylate side-chain of Asp145 and the neighboring Phe146 side-chain thus block the active site, thereby inactivating the enzyme. In the crystal structure of rHPAP(Pp), the enzyme "repression loop" has an open conformation similar to that observed in other mammalian PAP structures. The present structures demonstrate that the repression loop exhibits significant conformational flexibility, and the observed alternate binding mode suggests a possible inhibitory role for this loop.
Crystal structures of recombinant human purple Acid phosphatase with and without an inhibitory conformation of the repression loop.,Strater N, Jasper B, Scholte M, Krebs B, Duff AP, Langley DB, Han R, Averill BA, Freeman HC, Guss JM J Mol Biol. 2005 Aug 5;351(1):233-46. Epub 2005 Apr 26. PMID:15993892[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Briggs TA, Rice GI, Daly S, Urquhart J, Gornall H, Bader-Meunier B, Baskar K, Baskar S, Baudouin V, Beresford MW, Black GC, Dearman RJ, de Zegher F, Foster ES, Frances C, Hayman AR, Hilton E, Job-Deslandre C, Kulkarni ML, Le Merrer M, Linglart A, Lovell SC, Maurer K, Musset L, Navarro V, Picard C, Puel A, Rieux-Laucat F, Roifman CM, Scholl-Burgi S, Smith N, Szynkiewicz M, Wiedeman A, Wouters C, Zeef LA, Casanova JL, Elkon KB, Janckila A, Lebon P, Crow YJ. Tartrate-resistant acid phosphatase deficiency causes a bone dysplasia with autoimmunity and a type I interferon expression signature. Nat Genet. 2011 Feb;43(2):127-31. doi: 10.1038/ng.748. Epub 2011 Jan 9. PMID:21217755 doi:10.1038/ng.748
- ↑ Lausch E, Janecke A, Bros M, Trojandt S, Alanay Y, De Laet C, Hubner CA, Meinecke P, Nishimura G, Matsuo M, Hirano Y, Tenoutasse S, Kiss A, Rosa RF, Unger SL, Renella R, Bonafe L, Spranger J, Unger S, Zabel B, Superti-Furga A. Genetic deficiency of tartrate-resistant acid phosphatase associated with skeletal dysplasia, cerebral calcifications and autoimmunity. Nat Genet. 2011 Feb;43(2):132-7. doi: 10.1038/ng.749. Epub 2011 Jan 9. PMID:21217752 doi:10.1038/ng.749
- ↑ Strater N, Jasper B, Scholte M, Krebs B, Duff AP, Langley DB, Han R, Averill BA, Freeman HC, Guss JM. Crystal structures of recombinant human purple Acid phosphatase with and without an inhibitory conformation of the repression loop. J Mol Biol. 2005 Aug 5;351(1):233-46. Epub 2005 Apr 26. PMID:15993892 doi:10.1016/j.jmb.2005.04.014
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