A beta-turn rich barley seed protein is correctly folded in Escherichia coli.
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.
Tamas
L
L
Department of Agricultural Sciences, University of Bristol, United Kingdom.
Greenfield
J
J
Halford
N G
NG
Tatham
A S
AS
Shewry
P R
PR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003545:Cysteine / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005983:Glutens; D001467:Hordeum; D008969:Molecular Sequence Data; D009154:Mutation; D010940:Plant Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012542:Scattering, Radiation; D017421:Sequence Analysis; D014965:X-Rays
D010940:Plant Proteins; D011994:Recombinant Proteins; D005983:Glutens; D003545:Cysteine