title: Purification of an active species of recombinant kappa-bungarotoxin.
abstract: The expression of kappa-bungarotoxin in Escherichia coli from a synthetic gene results in the production of multiple species of polypeptide. These include not only biologically active kappa-bungarotoxin but also a variety of inactive species, which include inactive monomers as well as disulfide-linked polymeric species. Identification of these species and their separation from the biologically active recombinant toxin is necessary for the use of the toxin in physiological and biochemical studies. This has been accomplished by a combination of ion-exchange and reverse-phase chromatography which results in a homogeneous toxin preparation. The active material produced is sufficient for many types of biological studies and for mutagenesis experiments directed at determining the structure function relationships of toxin interactions with the neuronal nicotinic acetylcholine receptor. In addition, the kappa-bungarotoxin produced in this manner has the distinct advantage over venom-purified kappa-bungarotoxin of not being contaminated with other venom components which could potentially affect experimental observations.
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: J J
authors.initials: JJ
authors.lastname: Fiordalisi
authors.forename: R L
authors.initials: RL
authors.lastname: Gigowski
authors.forename: G A
authors.initials: GA
authors.lastname: Grant
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D002038:Bungarotoxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis; D012911:Snakes; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000596:Amino Acids; D002038:Bungarotoxins; D010446:Peptide Fragments; D011994:Recombinant Proteins