title: High-throughput sequence determination of cyclic peptide library members by partial Edman degradation/mass spectrometry.
abstract: Cyclic peptides provide attractive lead compounds for drug discovery and excellent molecular probes in biomedical research. Large combinatorial libraries of cyclic peptides can now be routinely synthesized by the split-and-pool method and screened against biological targets. However, post-screening sequence determination of hit peptides has been problematic. In this report, a high-throughput method for the sequence determination of cyclic peptide library members has been developed. TentaGel microbeads (90 mum) were spatially segregated into outer and inner layers; cyclic peptides were displayed on the bead surface, whereas the inner core of each bead contained the corresponding linear peptide as the encoding sequence. After screening of the cyclic peptide library against a macromolecular target, the identity of hit peptides was determined by sequencing the linear encoding peptides inside the bead using a partial Edman degradation/mass spectrometry method. On-bead screening of an octapeptide library (theoretical diversity of 160 000) identified cyclic peptides that bind to streptavidin. A 400-member library of tyrocidine A analogues was synthesized on TentaGel macrobeads and solution-phase screening of the library directly against bacterial cells identified a tyrocidine analogue of improved antibacterial activity. Our results demonstrate that the new method for cyclic peptide sequence determination is reliable, operationally simple, rapid, and inexpensive and should greatly expand the utility of cyclic peptides in biomedical research.
authors.affiliation: Department of Chemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, USA.
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Joo
authors.forename: Qing
authors.initials: Q
authors.lastname: Xiao
authors.forename: Yun
authors.initials: Y
authors.lastname: Ling
authors.forename: Bhaskar
authors.initials: B
authors.lastname: Gopishetty
authors.forename: Dehua
authors.initials: D
authors.lastname: Pei
mesh_terms: D000890:Anti-Infective Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000032:analysis* / Q000737:chemistry; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D019809:Streptavidin / Q000737:chemistry; D014440:Tyrocidine / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D009842:Oligopeptides; D019151:Peptide Library; D010456:Peptides, Cyclic; D014440:Tyrocidine; D019809:Streptavidin