One-step purification of a model periplasmic protein from inclusion bodies by its fusion to an effective metal-binding peptide.
It has been demonstrated that the addition of a metal-binding amino acid sequence to an exposed terminus of a protein can be useful for purification using immobilized metal affinity chromatography (IMAC). Polyhistidine extensions, wherein sequential histidyl residues are placed at the end of a protein, have been utilized for protein purification through IMAC. Natural metal-binding peptides may also serve as starting points for the design of an affinity tail. As a model system, an octapeptide derived from angiotensin I was fused to TEM-beta-lactamase. When the modified protein was expressed in Escherichia coli, a one-step purification of this recombinant protein was accomplished from resolubilized inclusion body material.
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Department of Chemical Engineering, University of Pittsburgh, Pennsylvania 15261.
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D000595:Amino Acid Sequence; D000803:Angiotensin I / Q000737:chemistry; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000302:isolation & purification*
D008670:Metals; D009842:Oligopeptides; D011994:Recombinant Proteins; D000803:Angiotensin I; D001616:beta-Galactosidase