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spinqualitylabelsall models 2fo5, resolution 2.200Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

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 SO4 and ACE 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

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