title: Substrate specificity of CDC2 kinase from human HeLa cells as determined with synthetic peptides and molecular modeling.
abstract: A systematic study was undertaken in order to assess the substrate specificity of cyclin-B/cell division control protein kinase (CDC2) isolated from human HeLa cells, using 13-15 residue peptides with a central histone-like KKSPKK motif as a model. Replacement of the proline residue by any of the other 19 amino acids or D-proline drastically reduces or abolishes phosphorylation by CDC2. Changing the basic residues to Ala on either side of the -SP- structure differentially reduces phosphorylation. Molecular modeling and dynamics simulation indicated that the phosphorylation site of the peptide may have to adopt a turn-like conformation that will orientate the charged and hydrophobic residues so as to allow interaction with postulated binding surfaces within the CDC2 active site. It thus appears that, of the 20 coded amino acids, only proline can provide this conformation in short peptides. This is in agreement with the finding that sarcosine can replace proline in this respect (S. Ando et al. Biochem. Biophys. Res. Commun. 195, 837-843, 1993).
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: R J
authors.initials: RJ
authors.lastname: Sanchez
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Guarnaccia
authors.forename: A
authors.initials: A
authors.lastname: Tossi
authors.forename: S
authors.initials: S
authors.lastname: Zahariev
authors.forename: S
authors.initials: S
authors.lastname: Pongor
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D016203:CDC2 Protein Kinase