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PDBsum entry 2fo5
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Hydrolase/hydrolase inhibitor
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PDB id
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2fo5
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References listed in PDB file
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Key reference
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Title
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Heterologous expression, Purification, Refolding, And structural-Functional characterization of ep-B2, A self-Activating barley cysteine endoprotease.
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Authors
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M.T.Bethune,
P.Strop,
Y.Tang,
L.M.Sollid,
C.Khosla.
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Ref.
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Chem Biol, 2006,
13,
637-647.
[DOI no: ]
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PubMed id
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Abstract
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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.
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Figure 2.
Figure 2. Crystal Structure of Mature EP-B2 Complexed with
Leupeptin
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Figure 6.
Figure 6. Activity of EP-B2 against the Proteolytically
Resistant 33-mer Peptide from α2-Gliadin
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The above figures are
reprinted
by permission from Cell Press:
Chem Biol
(2006,
13,
637-647)
copyright 2006.
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