title: Permuteins of interleukin 1 beta--a simplified approach for the construction of permutated proteins having new termini.
abstract: A technique for the rapid and simple generation of permutated versions of the interleukin-1 beta (IL-1 beta) gene is described. In this method, the human IL-1 beta cDNA is twice amplified by the polymerase chain reaction (PCR) and the resulting DNA fragments are ligated in tandem. Between the two genes, the DNA sequence encodes a short four amino acid loop to link the native N- and C-terminal ends of the IL-1 beta protein. By using PCR amplification from this starting template, a new version of the IL-1 beta cDNA was obtained that encodes a permutated form of the IL-1 beta protein where the new N- and C-terminal amino acids correspond to residues 65 and 64 of the native IL-1 beta sequence, respectively. The name 'permutein' is proposed to describe proteins generated by this technology. The molecular profile (IL-1 receptor binding, biologic activity and solution properties) of the IL-1 permutein produced by this technology, permutein 65/64, is shown to be identical to that of native IL-1 beta. The approach should be useful to define further the structural features of this protein that are important for its function.
authors.affiliation: Du Pont Merck Pharmaceutical Co., Wilmington, DE 19880-0400.
authors.forename: R A
authors.initials: RA
authors.lastname: Horlick
authors.forename: H J
authors.initials: HJ
authors.lastname: George
authors.forename: G M
authors.initials: GM
authors.lastname: Cooke
authors.forename: R J
authors.initials: RJ
authors.lastname: Tritch
authors.forename: R C
authors.initials: RC
authors.lastname: Newton
authors.forename: A
authors.initials: A
authors.lastname: Dwivedi
authors.forename: M
authors.initials: M
authors.lastname: Lischwe
authors.forename: F R
authors.initials: FR
authors.lastname: Salemme
authors.forename: P C
authors.initials: PC
authors.lastname: Weber
authors.forename: R
authors.initials: R
authors.lastname: Horuk
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D011869:Radioligand Assay; D011994:Recombinant Proteins / Q000138:chemical synthesis
chemical_list: D007375:Interleukin-1; D011994:Recombinant Proteins