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| | ==Crystal structure of purine nucleoside phosphorylase from Entamoeba histolytica== | | ==Crystal structure of purine nucleoside phosphorylase from Entamoeba histolytica== |
| - | <StructureSection load='3tl6' size='340' side='right' caption='[[3tl6]], [[Resolution|resolution]] 2.65Å' scene=''> | + | <StructureSection load='3tl6' size='340' side='right'caption='[[3tl6]], [[Resolution|resolution]] 2.65Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3tl6]] is a 6 chain structure with sequence from [http://en.wikipedia.org/wiki/Entamoeba_histolytica Entamoeba histolytica]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3TL6 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3TL6 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3tl6]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Entamoeba_histolytica Entamoeba histolytica]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3TL6 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3TL6 FirstGlance]. <br> |
| - | </td></tr><tr><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene><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]] 2.65Å</td></tr> |
| - | <tr><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">EHI_130960 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=5759 Entamoeba histolytica])</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><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Purine-nucleoside_phosphorylase Purine-nucleoside phosphorylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.2.1 2.4.2.1] </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=3tl6 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3tl6 OCA], [https://pdbe.org/3tl6 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3tl6 RCSB], [https://www.ebi.ac.uk/pdbsum/3tl6 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3tl6 ProSAT]</span></td></tr> |
| - | <tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3tl6 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3tl6 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3tl6 RCSB], [http://www.ebi.ac.uk/pdbsum/3tl6 PDBsum]</span></td></tr> | + | </table> |
| - | <table> | + | == Function == |
| - | <div style="background-color:#fffaf0;">
| + | [https://www.uniprot.org/uniprot/C4LXG4_ENTH1 C4LXG4_ENTH1] |
| - | == Publication Abstract from PubMed == | + | |
| - | Despite recent advances, the expression of heterologous proteins in Escherichia coli for crystallization remains a nontrivial challenge. The present study investigates the efficacy of maltose-binding protein (MBP) fusion as a general strategy for rescuing the expression of target proteins. From a group of sequence-verified clones with undetectable levels of protein expression in an E. coli T7 expression system, 95 clones representing 16 phylogenetically diverse organisms were selected for recloning into a chimeric expression vector with an N-terminal histidine-tagged MBP. PCR-amplified inserts were annealed into an identical ligation-independent cloning region in an MBP-fusion vector and were analyzed for expression and solubility by high-throughput nickel-affinity binding. This approach yielded detectable expression of 72% of the clones; soluble expression was visible in 62%. However, the solubility of most proteins was marginal to poor upon cleavage of the MBP tag. This study offers large-scale evidence that MBP can improve the soluble expression of previously non-expressing proteins from a variety of eukaryotic and prokaryotic organisms. While the behavior of the cleaved proteins was disappointing, further refinements in MBP tagging may permit the more widespread use of MBP-fusion proteins in crystallographic studies.
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| - | Expression of proteins in Escherichia coli as fusions with maltose-binding protein to rescue non-expressed targets in a high-throughput protein-expression and purification pipeline.,Hewitt SN, Choi R, Kelley A, Crowther GJ, Napuli AJ, Van Voorhis WC Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt, 9):1006-9. Epub 2011 Aug 13. PMID:21904041<ref>PMID:21904041</ref>
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| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
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| | ==See Also== | | ==See Also== |
| - | *[[Purine nucleoside phosphorylase|Purine nucleoside phosphorylase]] | + | *[[Purine nucleoside phosphorylase 3D structures|Purine nucleoside phosphorylase 3D structures]] |
| - | == References ==
| + | |
| - | <references/>
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| | [[Category: Entamoeba histolytica]] | | [[Category: Entamoeba histolytica]] |
| - | [[Category: Purine-nucleoside phosphorylase]] | + | [[Category: Large Structures]] |
| - | [[Category: Clifton, M C.]] | + | [[Category: Clifton MC]] |
| - | [[Category: Edwards, T E.]] | + | [[Category: Edwards TE]] |
| - | [[Category: SSGCID, Seattle Structural Genomics Center for Infectious Disease.]]
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| - | [[Category: Anaerobic parasitic protozoan]]
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| - | [[Category: Digestive tract cyst]]
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| - | [[Category: Fusion]]
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| - | [[Category: Maltose binding protein]]
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| - | [[Category: Mbp]]
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| - | [[Category: Nucleotide salvage pathway]]
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| - | [[Category: Pnpase]]
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| - | [[Category: Purine metabolism]]
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| - | [[Category: Seattle structural genomics center for infectious disease]]
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| - | [[Category: Ssgcid]]
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| - | [[Category: Structural genomic]]
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| - | [[Category: Transferase]]
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