Abstract
Wild-type and cysteine-containing mutant C hordeins from barley were expressed in Escherichia coli at high levels (> or = 30mg/liter). N-terminal sequence analysis, SDS-PAGE, RP-HPLC, cd spectroscopy, and small angle X-ray scattering demonstrated that their physicochemical properties were similar to those of C hordeins isolated from barley grain. This indicates that the expressed proteins were correctly folded. The cysteine-containing mutant showed evidence of polymer formation in E. coli, nonreduced preparations of the protein showing the presence of polymers that were replaced by a single protein when a reducing agent was added.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Chromatography, High Pressure Liquid
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Circular Dichroism
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Cysteine / genetics
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Escherichia coli / genetics
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Glutens
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Hordeum*
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Molecular Sequence Data
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Mutation
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Plant Proteins / biosynthesis
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Plant Proteins / chemistry*
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Plant Proteins / genetics
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Plant Proteins / isolation & purification
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Protein Folding*
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Protein Structure, Secondary*
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Scattering, Radiation
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Sequence Analysis
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X-Rays
Substances
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Plant Proteins
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Recombinant Proteins
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Glutens
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Cysteine