title: Phosphorylation effects on flanking charged residues. Structural implications for signal transduction in protein kinases.
abstract: 1H-NMR and 31P-NMR spectroscopy were employed to assess the electrostatic consequences of phosphorylation of single and multiple tyrosine residues in peptides derived from the core and tail autophosphorylation regions of the human insulin receptor tyrosine-kinase domain. In both peptides, phosphorylation was accompanied by changes in the resonances from basic side-chains; those from acidic residues were unaffected. Tyrosine phosphorylation caused increases of up to one in the pKa values of histidine residues situated up to eight residues away in the primary sequence. Titration curve analysis by Hill plots suggested some cooperativity of histidine and phosphate ionizations. Behaviour closely analogous to that of the insulin receptor tail peptide was observed during changes in phosphorylation of the intact insulin receptor kinase domain, suggesting that the electrostatic dissemination effects seen for the isolated peptide are retained by the peptide sequence in the context of the much larger protein. Similar changes in the behaviour of basic residues were also observed upon tyrosine phosphorylation of a cdc2-derived peptide, suggesting that this potential of phosphorylation events to propagate directed structural changes may find a widespread utility in the activation of protein kinases and in the transduction of phosphorylation-based signalling.
authors.affiliation: School of Biochemistry, University of Birmingham, England.
authors.forename: A
authors.initials: A
authors.lastname: Chavanieu
authors.forename: N E
authors.initials: NE
authors.lastname: Keane
authors.forename: P G
authors.initials: PG
authors.lastname: Quirk
authors.forename: B A
authors.initials: BA
authors.lastname: Levine
authors.forename: B
authors.initials: B
authors.lastname: Calas
authors.forename: L
authors.initials: L
authors.lastname: Wei
authors.forename: L
authors.initials: L
authors.lastname: Ellis
mesh_terms: D000595:Amino Acid Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011972:Receptor, Insulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D015398:Signal Transduction / Q000235:genetics*; D014443:Tyrosine / Q000378:metabolism
chemical_list: D010446:Peptide Fragments; D014443:Tyrosine; D006639:Histidine; D011972:Receptor, Insulin; D016203:CDC2 Protein Kinase