title: Biological significance of divalent metal ion binding to 14-3-3 proteins in relationship to nitrate reductase inactivation.
abstract: In this report we address two questions regarding the regulation of phosphorylated nitrate reductase (pNR; EC 1.6.6.1) by 14-3-3 proteins. The first concerns the requirement for millimolar concentrations of a divalent cation in order to form the inactive pNR:14-3-3 complex at pH 7.5. The second concerns the reduced requirement for divalent cations at pH 6.5. In answering these questions we highlight a possible general mechanism involved in the regulation of 14-3-3 binding to target proteins. We show that divalent cations (e.g. Ca2+, Mg2+ and Mn2+) bind directly to 14-3-3s, and as a result cause a conformational change, manifested as an increase in surface hydrophobicity. A similar change is also obtained by decreasing the pH from pH 7.5 to pH 6.5, in the absence of divalent cations, and we propose that protonation of amino acid residues brings about a similar effect to metal ion binding. A possible regulatory mechanism, where the 14-3-3 protein has to be "primed" prior to binding a target protein, is discussed.
authors.affiliation: US Department of Agriculture, Agricultural Research Service, Raleigh, NC 27695-7695, USA.
authors.forename: G S
authors.initials: GS
authors.lastname: Athwal
authors.forename: J L
authors.initials: JL
authors.lastname: Huber
authors.forename: S C
authors.initials: SC
authors.lastname: Huber
mesh_terms: D048948:14-3-3 Proteins; D001665:Binding Sites; D002413:Cations, Divalent; D008670:Metals / Q000378:metabolism*; D050895:Nitrate Reductase (NADH); D009565:Nitrate Reductases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010748:Phosphopeptides / Q000138:chemical synthesis / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D011522:Protons; D014446:Tyrosine 3-Monooxygenase
chemical_list: D048948:14-3-3 Proteins; D002413:Cations, Divalent; D008670:Metals; D010748:Phosphopeptides; D011506:Proteins; D011522:Protons; D014446:Tyrosine 3-Monooxygenase; D009565:Nitrate Reductases; D050895:Nitrate Reductase (NADH)