This old version of Proteopedia is provided for student assignments while the new version is undergoing repairs. Content and edits done in this old version of Proteopedia after March 1, 2026 will eventually be lost when it is retired in about June of 2026.

Apply for new accounts at the new Proteopedia. Your logins will work in both the old and new versions.


2fo5

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
(New page: 200px<br /><applet load="2fo5" size="450" color="white" frame="true" align="right" spinBox="true" caption="2fo5, resolution 2.200&Aring;" /> '''Crystal structure o...)
Current revision (15:30, 3 March 2021) (edit) (undo)
 
(13 intermediate revisions not shown.)
Line 1: Line 1:
-
[[Image:2fo5.gif|left|200px]]<br /><applet load="2fo5" size="450" color="white" frame="true" align="right" spinBox="true"
 
-
caption="2fo5, resolution 2.200&Aring;" />
 
-
'''Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin'''<br />
 
-
==Overview==
+
==Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin==
-
We describe the heterologous expression in Escherichia coli of the, proenzyme precursor to EP-B2, a cysteine endoprotease from germinating, barley seeds. High yields (50 mg/l) of recombinant proEP-B2 were obtained, from E. coli inclusion bodies in shake flask cultures following, purification and refolding. The zymogen was rapidly autoactivated to its, mature form under acidic conditions at a rate independent of proEP-B2, concentration, suggesting a cis mechanism of autoactivation. Mature EP-B2, was stable and active over a wide pH range and efficiently hydrolyzed a, recombinant wheat gluten protein, alpha2-gliadin, at sequences with known, immunotoxicity in celiac sprue patients. The X-ray crystal structure of, mature EP-B2 bound to leupeptin was solved to 2.2 A resolution and, provided atomic insights into the observed subsite specificity of the, endoprotease. Our findings suggest that orally administered proEP-B2 may, be especially well suited for treatment of celiac sprue.
+
<StructureSection load='2fo5' size='340' side='right'caption='[[2fo5]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
 +
== Structural highlights ==
 +
<table><tr><td colspan='2'>[[2fo5]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Barley Barley]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2FO5 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2FO5 FirstGlance]. <br>
 +
</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='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACE:ACETYL+GROUP'>ACE</scene>, <scene name='pdbligand=AR7:AMINO{[(4S)-4-AMINO-5,5-DIHYDROXYPENTYL]AMINO}METHANIMINIUM'>AR7</scene></td></tr>
 +
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">EPB2 ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=4513 Barley])</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=2fo5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2fo5 OCA], [https://pdbe.org/2fo5 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2fo5 RCSB], [https://www.ebi.ac.uk/pdbsum/2fo5 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2fo5 ProSAT]</span></td></tr>
 +
</table>
 +
== Evolutionary Conservation ==
 +
[[Image:Consurf_key_small.gif|200px|right]]
 +
Check<jmol>
 +
<jmolCheckbox>
 +
<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/fo/2fo5_consurf.spt"</scriptWhenChecked>
 +
<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
 +
<text>to colour the structure by Evolutionary Conservation</text>
 +
</jmolCheckbox>
 +
</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=2fo5 ConSurf].
 +
<div style="clear:both"></div>
 +
<div style="background-color:#fffaf0;">
 +
== Publication Abstract from PubMed ==
 +
We describe the heterologous expression in Escherichia coli of the proenzyme precursor to EP-B2, a cysteine endoprotease from germinating barley seeds. High yields (50 mg/l) of recombinant proEP-B2 were obtained from E. coli inclusion bodies in shake flask cultures following purification and refolding. The zymogen was rapidly autoactivated to its mature form under acidic conditions at a rate independent of proEP-B2 concentration, suggesting a cis mechanism of autoactivation. Mature EP-B2 was stable and active over a wide pH range and efficiently hydrolyzed a recombinant wheat gluten protein, alpha2-gliadin, at sequences with known immunotoxicity in celiac sprue patients. The X-ray crystal structure of mature EP-B2 bound to leupeptin was solved to 2.2 A resolution and provided atomic insights into the observed subsite specificity of the endoprotease. Our findings suggest that orally administered proEP-B2 may be especially well suited for treatment of celiac sprue.
-
==About this Structure==
+
Heterologous expression, purification, refolding, and structural-functional characterization of EP-B2, a self-activating barley cysteine endoprotease.,Bethune MT, Strop P, Tang Y, Sollid LM, Khosla C Chem Biol. 2006 Jun;13(6):637-47. PMID:16793521<ref>PMID:16793521</ref>
-
2FO5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Hordeum_vulgare Hordeum vulgare] with SO4 and ACE as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2FO5 OCA].
+
-
==Reference==
+
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
-
Heterologous expression, purification, refolding, and structural-functional characterization of EP-B2, a self-activating barley cysteine endoprotease., Bethune MT, Strop P, Tang Y, Sollid LM, Khosla C, Chem Biol. 2006 Jun;13(6):637-47. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16793521 16793521]
+
</div>
-
[[Category: Hordeum vulgare]]
+
<div class="pdbe-citations 2fo5" style="background-color:#fffaf0;"></div>
-
[[Category: Single protein]]
+
-
[[Category: Bethune, M.T.]]
+
-
[[Category: Brunger, A.T.]]
+
-
[[Category: Khosla, C.]]
+
-
[[Category: Strop, P.]]
+
-
[[Category: ACE]]
+
-
[[Category: SO4]]
+
-
[[Category: cysteine endoprotease]]
+
-
[[Category: endopeptidase]]
+
-
[[Category: ep-b2]]
+
-
[[Category: epb]]
+
-
[[Category: epb2]]
+
-
[[Category: leupeptin]]
+
-
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 10:39:11 2007''
+
==See Also==
 +
*[[Proteinase|Proteinase]]
 +
*[[Proteinase 3D structures|Proteinase 3D structures]]
 +
== References ==
 +
<references/>
 +
__TOC__
 +
</StructureSection>
 +
[[Category: Barley]]
 +
[[Category: Large Structures]]
 +
[[Category: Bethune, M T]]
 +
[[Category: Brunger, A T]]
 +
[[Category: Khosla, C]]
 +
[[Category: Strop, P]]
 +
[[Category: Cysteine endoprotease]]
 +
[[Category: Endopeptidase]]
 +
[[Category: Ep-b2]]
 +
[[Category: Epb]]
 +
[[Category: Epb2]]
 +
[[Category: Hydrolase]]
 +
[[Category: Hydrolase-hydrolase inhibitor complex]]
 +
[[Category: Leupeptin]]

Current revision

Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin

PDB ID 2fo5

Drag the structure with the mouse to rotate

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/2fo5"
Personal tools