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==Structural Studies and Protein Engineering of Inositol Phosphate Multikinase==
==Structural Studies and Protein Engineering of Inositol Phosphate Multikinase==
<StructureSection load='4frf' size='340' side='right' caption='[[4frf]], [[Resolution|resolution]] 2.90&Aring;' scene=''>
<StructureSection load='4frf' size='340' side='right'caption='[[4frf]], [[Resolution|resolution]] 2.90&Aring;' scene=''>
== Structural highlights ==
== Structural highlights ==
<table><tr><td colspan='2'>[[4frf]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Arath Arath]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4FRF OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4FRF FirstGlance]. <br>
<table><tr><td colspan='2'>[[4frf]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Arabidopsis_thaliana Arabidopsis thaliana]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4FRF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4FRF FirstGlance]. <br>
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></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]] 2.9&#8491;</td></tr>
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">At5g07370, AtIpmk alpha, IP3K, IPK2a, IPMK, T2I1.80 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=3702 ARATH])</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></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=4frf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4frf OCA], [http://pdbe.org/4frf PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4frf RCSB], [http://www.ebi.ac.uk/pdbsum/4frf PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4frf ProSAT]</span></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=4frf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4frf OCA], [https://pdbe.org/4frf PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4frf RCSB], [https://www.ebi.ac.uk/pdbsum/4frf PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4frf ProSAT]</span></td></tr>
</table>
</table>
== Function ==
== Function ==
[[http://www.uniprot.org/uniprot/IPMKA_ARATH IPMKA_ARATH]] Inositol phosphate kinase with a broad substrate specificity. Phosphorylates inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), inositol 1,4,5,6-tetrakisphosphate (Ins(1,4,5,6)P4), inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4), inositol 1,3,4,6-tetrakisphosphate (Ins(1,3,4,6)P4) and inositol 1,2,3,4,6-pentakisphosphate (Ins(1,2,3,4,6)P5) but not inositol 1,4-bisphosphate (Ins(1,4)P2), inositol 1,3,4-trisphosphate (Ins(1,3,4)P3), inositol 1,2,6-trisphosphate (Ins(1,2,6)P3), inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P4), inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5), inositol 1,2,4,5,6-pentakisphosphate (Ins(1,2,4,5,6)P5) or inositol hexakisphosphate (InsP6). Regulates pollen and root development probably through the regulation of InsP3-mediated calcium accumulation.<ref>PMID:12226109</ref> <ref>PMID:15618435</ref> 
[https://www.uniprot.org/uniprot/IPMKA_ARATH IPMKA_ARATH] Inositol phosphate kinase with a broad substrate specificity. Phosphorylates inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), inositol 1,4,5,6-tetrakisphosphate (Ins(1,4,5,6)P4), inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4), inositol 1,3,4,6-tetrakisphosphate (Ins(1,3,4,6)P4) and inositol 1,2,3,4,6-pentakisphosphate (Ins(1,2,3,4,6)P5) but not inositol 1,4-bisphosphate (Ins(1,4)P2), inositol 1,3,4-trisphosphate (Ins(1,3,4)P3), inositol 1,2,6-trisphosphate (Ins(1,2,6)P3), inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P4), inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5), inositol 1,2,4,5,6-pentakisphosphate (Ins(1,2,4,5,6)P5) or inositol hexakisphosphate (InsP6). Regulates pollen and root development probably through the regulation of InsP3-mediated calcium accumulation.<ref>PMID:12226109</ref> <ref>PMID:15618435</ref>  
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Inositol phosphates (IPs) regulate vital processes in eukaryotes, and their production downstream of phospholipase C activation is controlled through a network of evolutionarily conserved kinases and phosphatases. Inositol phosphate multikinase (IPMK, also called Ipk2 and Arg82) accounts for phosphorylation of IP(3) to IP(5), as well as production of several other IP molecules. Here, we report the structure of Arabidopsis thaliana IPMKalpha at 2.9 A and find it is similar to the yeast homolog Ipk2, despite 17% sequence identity, as well as the active site architecture of human IP(3) 3-kinase. Structural comparison and substrate modeling were used to identify a putative basis for IPMK selectivity. To test this model, we re-engineered binding site residues predicted to have restricted substrate specificity. Using steady-state kinetics and in vivo metabolic labeling studies in modified yeast strains, we observed that K117W and K117W:K121W mutants exhibited nearly normal 6-kinase function but harbored significantly reduced 3-kinase activity. These mutants complemented conditional nutritional growth defects observed in ipmk null yeast and, remarkably, suppressed lethality observed in ipmk null flies. Our data are consistent with the hypothesis that IPMK 6-kinase activity and production of Ins(1,4,5,6)P(4) are critical for cellular signaling. Overall, our studies provide new insights into the structure and function of IPMK and utilize a synthetic biological approach to redesign inositol phosphate signaling pathways.
 
Structural studies and protein engineering of inositol phosphate multikinase.,Endo-Streeter S, Tsui MK, Odom AR, Block J, York JD J Biol Chem. 2012 Oct 12;287(42):35360-9. doi: 10.1074/jbc.M112.365031. Epub 2012, Aug 15. PMID:22896696<ref>PMID:22896696</ref>
 
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 4frf" style="background-color:#fffaf0;"></div>
== References ==
== References ==
<references/>
<references/>
__TOC__
__TOC__
</StructureSection>
</StructureSection>
[[Category: Arath]]
[[Category: Arabidopsis thaliana]]
[[Category: Endo-Streeter, S T]]
[[Category: Large Structures]]
[[Category: Odom, A R]]
[[Category: Endo-Streeter ST]]
[[Category: Tsui, M]]
[[Category: Odom AR]]
[[Category: York, J D]]
[[Category: Tsui M]]
[[Category: Atp grasp]]
[[Category: York JD]]
[[Category: Inositol phosphate kinase]]
[[Category: Transferase]]

Latest revision as of 14:23, 1 March 2024

Structural Studies and Protein Engineering of Inositol Phosphate MultikinaseStructural Studies and Protein Engineering of Inositol Phosphate Multikinase

Structural highlights

4frf is a 2 chain structure with sequence from Arabidopsis thaliana. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 2.9Å
Ligands:
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

IPMKA_ARATH Inositol phosphate kinase with a broad substrate specificity. Phosphorylates inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), inositol 1,4,5,6-tetrakisphosphate (Ins(1,4,5,6)P4), inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4), inositol 1,3,4,6-tetrakisphosphate (Ins(1,3,4,6)P4) and inositol 1,2,3,4,6-pentakisphosphate (Ins(1,2,3,4,6)P5) but not inositol 1,4-bisphosphate (Ins(1,4)P2), inositol 1,3,4-trisphosphate (Ins(1,3,4)P3), inositol 1,2,6-trisphosphate (Ins(1,2,6)P3), inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P4), inositol 1,3,4,5,6-pentakisphosphate (Ins(1,3,4,5,6)P5), inositol 1,2,4,5,6-pentakisphosphate (Ins(1,2,4,5,6)P5) or inositol hexakisphosphate (InsP6). Regulates pollen and root development probably through the regulation of InsP3-mediated calcium accumulation.[1] [2]

References

  1. ↑ Stevenson-Paulik J, Odom AR, York JD. Molecular and biochemical characterization of two plant inositol polyphosphate 6-/3-/5-kinases. J Biol Chem. 2002 Nov 8;277(45):42711-8. Epub 2002 Sep 10. PMID:12226109 doi:http://dx.doi.org/10.1074/jbc.M209112200
  2. ↑ Xu J, Brearley CA, Lin WH, Wang Y, Ye R, Mueller-Roeber B, Xu ZH, Xue HW. A role of Arabidopsis inositol polyphosphate kinase, AtIPK2alpha, in pollen germination and root growth. Plant Physiol. 2005 Jan;137(1):94-103. Epub 2004 Dec 23. PMID:15618435 doi:http://dx.doi.org/pp.104.045427

4frf, resolution 2.90Å

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