title: A CDC6 protein-binding peptide selected using a bacterial two-hybrid-like system is a cell cycle inhibitor.
abstract: Peptides or small molecules able to modulate protein-protein interactions hold promise as tools with which to probe and manipulate biological pathways. An important issue in this nascent field is to evaluate different methods with which to search libraries for molecules that modulate the function of specific target proteins. One strategy is to screen libraries for molecules that bind specifically to a protein known to be critical in the pathway of interest, with the expectation that the molecules isolated will recognize regions of the target protein important for its function and thereby exhibit biological activity. Here, a peptide library was screened using a two-hybrid-like system for molecules able to bind human CDC6 protein (CDC6p), required for the initiation of DNA replication in eukaryotic cells. From a collection of over a million peptides, a single species that exhibited good affinity and specificity for binding CDC6p was obtained. When expressed in human cells, the peptide inhibited cell cycle progression and exhibited other properties expected of a CDC6p inhibitor. This approach, which does not require detailed knowledge of the mechanism of action of a protein target, may be generally useful for isolating peptides capable of manipulating biological pathways.
authors.affiliation: Ryburn Center for Molecular Cardiology and Center for Biomedical Inventions, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-8573, USA.
authors.forename: W
authors.initials: W
authors.lastname: Zhu
authors.forename: R S
authors.initials: RS
authors.lastname: Williams
authors.forename: T
authors.initials: T
authors.lastname: Kodadek
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000235:genetics; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D002645:Chickens; D002843:Chromatin / Q000378:metabolism; D004261:DNA Replication; D016193:G1 Phase; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D016196:S Phase; D013379:Substrate Specificity; D014162:Transfection; D020798:Two-Hybrid System Techniques
chemical_list: C103706:CDC6 protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D009687:Nuclear Proteins; D019151:Peptide Library; D010455:Peptides