title: Design and synthesis of a peptide derived from positions 195-244 of human cdc25C phosphatase.
abstract: We have designed, synthesized and purified a 51 amino acid peptide derived from an essential domain of human cdc25C phosphatase. In vivo, differential phosphorylation of this domain regulates either the induction of mitotic processes, or the checkpoint arrest of eukaryotic cells in response to DNA damage. Peptide synthesis was achieved using the stepwise Fmoc strategy and resulted in an important yield of highly pure peptide. The final peptide was identified by amino acid analysis, electrospray mass spectrometry and nuclear magnetic resonance, which revealed that one of the two methionines within the peptide was oxidized into its sulphoxide derivative We investigated whether this 51 amino acid peptide folded into secondary structures in solution by circular dichroism and observed the formation of alpha helices in TFE. Finally, we verified that this peptide could bind to its biologically relevant 14-3-3 partner in vitro by fluorescence spectroscopy.
authors.affiliation: CRBM, CNRS-UPR 1086, Montpellier, France.
authors.forename: M C
authors.initials: MC
authors.lastname: Morris
authors.forename: J
authors.initials: J
authors.lastname: Mery
authors.forename: A
authors.initials: A
authors.lastname: Heitz
authors.forename: F
authors.initials: F
authors.lastname: Heitz
authors.forename: G
authors.initials: G
authors.lastname: Divita
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D020687:cdc25 Phosphatases
chemical_list: D018797:Cell Cycle Proteins; D010455:Peptides; D010749:Phosphoprotein Phosphatases; C517009:CDC25C protein, human; D020687:cdc25 Phosphatases