title: Effect of potassium ion on the phosphorus-31 nuclear magnetic resonance spectrum of the pyridoxal 5'-phosphate cofactor of Escherichia coli D-serine dehydratase.
abstract: 31P NMR studies were undertaken to determine how potassium ion increases the cofactor affinity of Escherichia coli D-serine dehydratase, a model pyridoxal 5'-phosphate requiring enzyme that converts the growth inhibitor D-serine to pyruvate and ammonia. Potassium ion was shown to promote the appearance of a second upfield shifted cofactor 31P resonance at 4.0 ppm (pH 7.8, 25 degrees C), that increased in area at the expense of the resonance at 4.4 ppm observed in the absence of K+. Na+ antagonized the K+ promoted appearance of the second resonance. These observations suggest that K+ and Na+ stabilize conformational states that differ with respect to O-P-O bond angle, conformation, and/or hydrogen bonding of the phosphate group. An analysis of the dependence of the relative intensities of the two resonances on the K+ concentration yielded a value of ca. 10 mM for the equilibrium constant for dissociation of K+ from D-serine dehydratase. The chemical shift difference between the two resonances indicated that the K+-stabilized and Na+-stabilized forms of the enzyme interconvert at a frequency less than 16 s-1 at pH 7.8, 25 degrees C.
authors.affiliation: Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
authors.forename: C L
authors.initials: CL
authors.lastname: Kojiro
authors.forename: M
authors.initials: M
authors.lastname: Marceau
authors.forename: J A
authors.initials: JA
authors.lastname: Shafer
mesh_terms: D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D012695:L-Serine Dehydratase / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010758:Phosphorus; D011188:Potassium / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology*; D011485:Protein Binding; D011732:Pyridoxal Phosphate / Q000378:metabolism*
chemical_list: D010758:Phosphorus; D011732:Pyridoxal Phosphate; D011189:Potassium Chloride; D012695:L-Serine Dehydratase; D011188:Potassium