Design and recognition properties of a hydropathically complementary peptide to human interleukin 1 beta.
A computer-designed hydropathically complementary peptide to human interleukin 1 beta (IL1 beta) precursor sequence 204-215 recognized the 204-215 peptide as well the entire IL1 beta protein with binding affinities in the micromolar range. Interaction between the complementary pair was characterized by analytical high-performance liquid affinity chromatography on columns derivatized with the computer-generated peptide. Recognition selectivity was clearly shown by the ability of the computer-generated complementary peptide columns to purify the IL1 beta-(204-215)-peptide from complex synthetic mixtures with high yields, independently of the type of solid support used. Recognition specificity was demonstrated by the inability of the IL1 beta-(204-215)-peptide and IL1 beta molecules to interact with blank columns or columns derivatized with other non-related peptides. Furthermore, scrambling the sequence of the computer-generated peptide or the IL1 beta-(204-215)-peptide in such a way as to alter their hydropathic profiles had the effect of abolishing binding. The complementary pair failed to interact in the presence of competing peptide, thus providing further evidence of specificity. Computer-generated complementary peptide affinity columns also proved useful for purification of recombinant human IL1 beta protein directly from crude Escherichia coli lysates.
Fassina
G
G
Protein Engineering Unit, TECNOGEN S.c.p.A., Milano, Italy.
Cassani
G
G
D000595:Amino Acid Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003201:Computers; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry* / Q000302:isolation & purification*; D053583:Interleukin-1beta; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification*
D007375:Interleukin-1; D053583:Interleukin-1beta; D010446:Peptide Fragments; C073692:interleukin-1beta (204-215)