3rbu: Difference between revisions

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[[Image:3rbu.png|left|200px]]


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==N-terminally AviTEV-tagged Human Glutamate Carboxypeptidase II in complex with 2-PMPA==
The line below this paragraph, containing "STRUCTURE_3rbu", creates the "Structure Box" on the page.
<StructureSection load='3rbu' size='340' side='right'caption='[[3rbu]], [[Resolution|resolution]] 1.60&Aring;' scene=''>
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== Structural highlights ==
or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
<table><tr><td colspan='2'>[[3rbu]] 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=3RBU OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3RBU FirstGlance]. <br>
or leave the SCENE parameter empty for the default display.
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.6&#8491;</td></tr>
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<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=BMA:BETA-D-MANNOSE'>BMA</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=G88:(2S)-2-(PHOSPHONOMETHYL)PENTANEDIOIC+ACID'>G88</scene>, <scene name='pdbligand=MAN:ALPHA-D-MANNOSE'>MAN</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
{{STRUCTURE_3rbu|  PDB=3rbu  |  SCENE=  }}
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=3rbu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3rbu OCA], [https://pdbe.org/3rbu PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3rbu RCSB], [https://www.ebi.ac.uk/pdbsum/3rbu PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3rbu ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/FOLH1_HUMAN FOLH1_HUMAN] Has both folate hydrolase and N-acetylated-alpha-linked-acidic dipeptidase (NAALADase) activity. Has a preference for tri-alpha-glutamate peptides. In the intestine, required for the uptake of folate. In the brain, modulates excitatory neurotransmission through the hydrolysis of the neuropeptide, N-aceylaspartylglutamate (NAAG), thereby releasing glutamate. Isoform PSM-4 and isoform PSM-5 would appear to be physiologically irrelevant. Involved in prostate tumor progression.  Also exhibits a dipeptidyl-peptidase IV type activity. In vitro, cleaves Gly-Pro-AMC.
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Affinity purification is a useful approach for purification of recombinant proteins. Eukaryotic expression systems have become more frequently used at the expense of prokaryotic systems since they afford recombinant eukaryotic proteins with post-translational modifications similar or identical to the native ones. Here, we present a one-step affinity purification set-up suitable for the purification of secreted proteins. The set-up is based on the interaction between biotin and mutated streptavidin. Drosophila Schneider 2 cells are chosen as the expression host, and a biotin acceptor peptide is used as an affinity tag. This tag is biotinylated by Escherichia coli biotin-protein ligase in vivo. We determined that localization of the ligase within the ER led to the most effective in vivo biotinylation of the secreted proteins. We optimized a protocol for large-scale expression and purification of AviTEV-tagged recombinant human glutamate carboxypeptidase II (Avi-GCPII) with milligram yields per liter of culture. We also determined the 3D structure of Avi-GCPII by X-ray crystallography and compared the enzymatic characteristics of the protein to those of its non-tagged variant. These experiments confirmed that AviTEV tag does not affect the biophysical properties of its fused partner. Purification approach, developed here, provides not only a sufficient amount of highly homogenous protein but also specifically and effectively biotinylates a target protein and thus enables its subsequent visualization or immobilization.


===N-terminally AviTEV-tagged Human Glutamate Carboxypeptidase II in complex with 2-PMPA===
Efficient and versatile one-step affinity purification of in vivo biotinylated proteins: Expression, characterization and structure analysis of recombinant human glutamate carboxypeptidase II.,Tykvart J, Sacha P, Barinka C, Knedlik T, Starkova J, Lubkowski J, Konvalinka J Protein Expr Purif. 2011 Dec 8. PMID:22178733<ref>PMID:22178733</ref>


From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 3rbu" style="background-color:#fffaf0;"></div>


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==See Also==
The line below this paragraph, {{ABSTRACT_PUBMED_22178733}}, adds the Publication Abstract to the page
*[[Carboxypeptidase 3D structures|Carboxypeptidase 3D structures]]
(as it appears on PubMed at http://www.pubmed.gov), where 22178733 is the PubMed ID number.
== References ==
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<references/>
{{ABSTRACT_PUBMED_22178733}}
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</StructureSection>
==About this Structure==
[[3rbu]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3RBU OCA].
 
==Reference==
<ref group="xtra">PMID:022178733</ref><references group="xtra"/>
[[Category: Glutamate carboxypeptidase II]]
[[Category: Homo sapiens]]
[[Category: Homo sapiens]]
[[Category: Barinka, C.]]
[[Category: Large Structures]]
[[Category: Knedlik, T.]]
[[Category: Barinka C]]
[[Category: Konvalinka, J.]]
[[Category: Knedlik T]]
[[Category: Lubkowski, J.]]
[[Category: Konvalinka J]]
[[Category: Sacha, P.]]
[[Category: Lubkowski J]]
[[Category: Starkova, J.]]
[[Category: Sacha P]]
[[Category: Tykvart, J.]]
[[Category: Starkova J]]
[[Category: Biotinylation]]
[[Category: Tykvart J]]
[[Category: Extracellular]]
[[Category: Hydrolase-hydrolase inhibitor complex]]
[[Category: Peptidase]]
Retrieved from "https://proteopedia.org/3rbu"