title: Trapping of peptide-based surrogates in an artificially created channel of cytochrome c peroxidase.
abstract: As recently described, the deliberate removal of the proposed electron transfer pathway from cytochrome c peroxidase resulted in the formation of an extended ligand-binding channel. The engineered channel formed a template for the removed peptide segment, suggesting that synthetic surrogates might be introduced to replace the native electron transfer pathway. This approach could be united with the recent development of sensitizer-linked substrates to initiate and study electron transfer, allowing access to unresolved issues about redox mechanism of the enzyme. Here, we present the design, synthesis, and screening of a peptide library containing natural and unnatural amino acids to identify the structural determinants for binding this channel mutant. Only one peptide, (benzimidazole-propionic acid)-Gly-Ala-Ala, appeared to interact, and gave evidence for both reversible and kinetically trapped binding, suggesting multiple conformations for the channel protein. Notably, this peptide was the most analogous to the removed electron transfer sequence, supporting the use of a cavity-template strategy for design of specific sensitizer-linked substrates as replacements for the native electron transfer pathway.
authors.affiliation: Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Anna-Maria A
authors.initials: AM
authors.lastname: Hays
authors.forename: Harry B
authors.initials: HB
authors.lastname: Gray
authors.forename: David B
authors.initials: DB
authors.lastname: Goodin
mesh_terms: D003578:Cytochrome-c Peroxidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004579:Electron Transport; D006860:Hydrogen Bonding; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D003578:Cytochrome-c Peroxidase