Complexes of Escherichia coli adenylate kinase and nucleotides: sup 1 H NMR studies of the nucleotide sites in solution

Journal Article · Tue Aug 14 00:00:00 EDT 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00484a015· OSTI ID:6058337

Vetter, I R; Reinstein, J; Roesch, P ^[1]

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1. Max Planck Institute for Medical Research, Heidelberg (West Germany)

One- and two-dimensional nuclear magnetic resonance (NMR) studies, in particular substrate-protein nuclear Overhauser effect (NOESY) measurements, as well as nucleotide and P{sup 1},P{sup 5}-bis-(5{prime}-adenosyl) pentaphosphate (AP{sub 5}A) titrations and studies of the temperature-dependent unfolding of the tertiary structure of Escherichia coli adenylate kinase (AK{sub EC}) were performed. These experiments and comparison with the same type of experiments performed with the porcine enzyme led them to the following conclusions: (1) at pH 8 and concentrations of approximately 2.5-3 mM, AK{sub EC} is partially unfolded at 318 K; (2) ATP{center dot}Mg{sup 2+} binds to the ATP site with a dissociation constant of approximately 40 {mu}M under the assumption that ATP binds to one nucleotide site only; (3) AP{sub 5}A{center dot}Mg{sup 2+} binds to both nucleotide sites and thus simulates the active complex; (4) the ATP{center dot}Mg{sup 2+} adenine in the AK{sub EC}{center dot}AP{sub 5}A{center dot}Mg{sup 2+} complex is located close to His{sup 134} and Phe{sup 19}; (5) the AK{sub EC} G-loop with bound ATP{center dot}Mg{sup 2+} is structurally highly homologous to the loop region in the oncogene product p21 with bound GTP{center dot}Mg{sup 2+}.

• OSTI ID: 6058337
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:32; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
AMINES
AROMATICS
AZAARENES
BACTERIA
BARYONS
ELEMENTARY PARTICLES
ENZYMES
ESCHERICHIA COLI
FERMIONS
GUANINE
HADRONS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
MAGNETIC RESONANCE
MICROORGANISMS
NUCLEAR MAGNETIC RESONANCE
NUCLEONS
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTONS
PURINES
RESONANCE
SUBSTRATES
TEMPERATURE DEPENDENCE
TRANSFERASES