4kjd: Difference between revisions
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==RatIntestinal AP expressed in E. coli== | ==RatIntestinal AP expressed in E. coli== | ||
<StructureSection load='4kjd' size='340' side='right' caption='[[4kjd]], [[Resolution|resolution]] 2.21Å' scene=''> | <StructureSection load='4kjd' size='340' side='right'caption='[[4kjd]], [[Resolution|resolution]] 2.21Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[4kjd]] is a 2 chain structure with sequence from [ | <table><tr><td colspan='2'>[[4kjd]] 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=4KJD OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4KJD FirstGlance]. <br> | ||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene> | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=SEP:PHOSPHOSERINE'>SEP</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=4kjd FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kjd OCA], [https://pdbe.org/4kjd PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4kjd RCSB], [https://www.ebi.ac.uk/pdbsum/4kjd PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4kjd ProSAT]</span></td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | |||
</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: | [[Category: Large Structures]] | ||
[[Category: | [[Category: Rattus norvegicus]] | ||
[[Category: Anumula | [[Category: Anumula RK]] | ||
[[Category: Ghosh | [[Category: Ghosh K]] | ||
[[Category: Laksmaiah | [[Category: Laksmaiah BK]] | ||
Revision as of 11:54, 7 December 2022
RatIntestinal AP expressed in E. coliRatIntestinal AP expressed in E. coli
Structural highlights
FunctionPublication Abstract from PubMedIntestinal 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 AlsoReferences
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