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| | ==Structure of Rat Intestinal Alkaline Phosphatase expressed in insect cell== | | ==Structure of Rat Intestinal Alkaline Phosphatase expressed in insect cell== |
| - | <StructureSection load='4kjg' size='340' side='right' caption='[[4kjg]], [[Resolution|resolution]] 2.38Å' scene=''> | + | <StructureSection load='4kjg' size='340' side='right'caption='[[4kjg]], [[Resolution|resolution]] 2.38Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4kjg]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Buffalo_rat Buffalo rat]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KJG OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4KJG FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4kjg]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KJG OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4KJG FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=4NP:4-NITROPHENYL+PHOSPHATE'>4NP</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=4NP:4-NITROPHENYL+PHOSPHATE'>4NP</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4kjd|4kjd]]</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=4kjg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kjg OCA], [https://pdbe.org/4kjg PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4kjg RCSB], [https://www.ebi.ac.uk/pdbsum/4kjg PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4kjg ProSAT]</span></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">Alpi ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=10116 Buffalo rat])</td></tr>
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| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Alkaline_phosphatase Alkaline phosphatase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.3.1 3.1.3.1] </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=4kjg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kjg OCA], [http://pdbe.org/4kjg PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4kjg RCSB], [http://www.ebi.ac.uk/pdbsum/4kjg PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4kjg ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/PPBI1_RAT PPBI1_RAT] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | ==See Also== | | ==See Also== |
| - | *[[Alkaline phosphatase|Alkaline phosphatase]] | + | *[[Alkaline phosphatase 3D structures|Alkaline phosphatase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Alkaline phosphatase]] | + | [[Category: Large Structures]] |
| - | [[Category: Buffalo rat]] | + | [[Category: Rattus norvegicus]] |
| - | [[Category: Anumula, R K]] | + | [[Category: Anumula RK]] |
| - | [[Category: Ghosh, K]] | + | [[Category: Ghosh K]] |
| - | [[Category: Laksmaiah, B K]] | + | [[Category: Laksmaiah BK]] |
| - | [[Category: Alpha/beta fold]]
| + | |
| - | [[Category: Hydrolase]]
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| Structural highlights
Function
PPBI1_RAT
Publication Abstract from PubMed
Intestinal alkaline phosphatases (IAPs) are involved in the cleavage of phosphate prodrugs to liberate the drug for absorption in the intestine. To facilitate in vitro characterization of phosphate prodrugs, we have cloned, expressed, purified and characterized IAPs from rat and cynomolgus monkey (rIAP and cIAP respectively) which are important pre-clinical species for drug metabolism studies. The recombinant rat and monkey enzymes expressed in Sf9 insect cells (IAP-Ic) were found to be glycosylated and active. Expression of rat IAP in Escherichia coli (rIAP-Ec) led to approximately 200-fold loss of activity that was partially recovered by the addition of external Zn2+ and Mg2+ ions. Crystal structures of rIAP-Ec and rIAP-Ic were determined and they provide rationale for the discrepancy in enzyme activities. Rat IAP-Ic retains its activity in presence of both Zn2+ and Mg2+ whereas activity of most other alkaline phosphatases (APs) including the cIAP was strongly inhibited by excess Zn2+. Based on our crystal structure, we hypothesized the residue Q317 in rIAP, present within 7A of the Mg2+ at M3, to be important for this difference in activity. The Q317H rIAP and H317Q cIAP mutants showed reversal in effect of Zn2+, corroborating the hypothesis. Further analysis of the two structures indicated a close linkage between glycosylation and crown domain stability. A triple mutant of rIAP, where all the three putative N-linked glycosylation sites were mutated showed thermal instability and reduced activity.
Crystal structure of rat intestinal alkaline phosphatase - Role of crown domain in mammalian alkaline phosphatases.,Ghosh K, Mazumder Tagore D, Anumula R, Lakshmaiah B, Pabbisetty K, Singaram S, Matan T, Kallipatti S, Selvam S, Krishnamurthy P, Ramarao M J Struct Biol. 2013 Sep 27. pii: S1047-8477(13)00258-X. doi:, 10.1016/j.jsb.2013.09.017. PMID:24076154[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Ghosh K, Mazumder Tagore D, Anumula R, Lakshmaiah B, Pabbisetty K, Singaram S, Matan T, Kallipatti S, Selvam S, Krishnamurthy P, Ramarao M. Crystal structure of rat intestinal alkaline phosphatase - Role of crown domain in mammalian alkaline phosphatases. J Struct Biol. 2013 Sep 27. pii: S1047-8477(13)00258-X. doi:, 10.1016/j.jsb.2013.09.017. PMID:24076154 doi:http://dx.doi.org/10.1016/j.jsb.2013.09.017
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