2fo5
From Proteopedia
(New page: 200px<br /><applet load="2fo5" size="450" color="white" frame="true" align="right" spinBox="true" caption="2fo5, resolution 2.200Å" /> '''Crystal structure o...) |
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| - | [[Image:2fo5.gif|left|200px]]<br /><applet load="2fo5" size=" | + | [[Image:2fo5.gif|left|200px]]<br /><applet load="2fo5" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2fo5, resolution 2.200Å" /> | caption="2fo5, resolution 2.200Å" /> | ||
'''Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin'''<br /> | '''Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin'''<br /> | ||
==Overview== | ==Overview== | ||
| - | We describe the heterologous expression in Escherichia coli of the | + | 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== | ==About this Structure== | ||
| - | 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:// | + | 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 <scene name='pdbligand=SO4:'>SO4</scene> and <scene name='pdbligand=ACE:'>ACE</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2FO5 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Hordeum vulgare]] | [[Category: Hordeum vulgare]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Bethune, M | + | [[Category: Bethune, M T.]] |
| - | [[Category: Brunger, A | + | [[Category: Brunger, A T.]] |
[[Category: Khosla, C.]] | [[Category: Khosla, C.]] | ||
[[Category: Strop, P.]] | [[Category: Strop, P.]] | ||
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[[Category: leupeptin]] | [[Category: leupeptin]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:23:22 2008'' |
Revision as of 15:23, 21 February 2008
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Crystal structure of recombinant barley cysteine endoprotease B isoform 2 (EP-B2) in complex with leupeptin
Overview
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
2FO5 is a Single protein structure of sequence from Hordeum vulgare with and as ligands. Full crystallographic information is available from OCA.
Reference
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
Page seeded by OCA on Thu Feb 21 17:23:22 2008
Categories: Hordeum vulgare | Single protein | Bethune, M T. | Brunger, A T. | Khosla, C. | Strop, P. | ACE | SO4 | Cysteine endoprotease | Endopeptidase | Ep-b2 | Epb | Epb2 | Leupeptin
