Complexes of yeast adenylate kinase and nucleotides investigated by sup 1 H NMR
- Max Planck Institute for Medical Research, Heidelberg (West Germany)
- Max Planck Institute for Biophysical Chemistry, Goettingen (West Germany)
- Univ. of Bayreuth (West Germany)
The role of one of the histidine residues present in many adenylate kinases (H36 in the porcine cytosolic enzyme) is highly disputed. The authors studied the yeast enzyme (AK{sub ye}) containing this His residue. AK{sub ye} is highly homologous to the Escherichia coli enzyme (AK{sub ec}), a protein that is already well characterized by NMR and does not contain the His residue in question. In addition, discrepancies between solution structural and X-ray crystallographic studies on the location of the nucleotide binding sites of adenylate kinases are clarified. One- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy was used to investigate AK{sub ye} and its complex with the bisubstrate analogue P{sup 1},P{sup 5}-bis(5{prime}-adenosyl)pentaphosphate (AP{sub 5}A). From these studies, all aromatic residues of AK{sub ec} involved in the binding of ATP{center dot}Mg{sup 2+} have functional analogues in AK{sub ye}. The AMP site seems to make no contacts to aromatic side chains, neither in the AK{sub ye}{center dot}AP{sub 5}A{center dot}Mg{sup 2+} nor in the AK{sub ec}{center dot}AP{sub 5}A{center dot}Mg{sup 2+} complexes, so that it is presently not possible to localize this binding site by NMR. In combination with the recent X-ray results on the AP{sub 5}A complexes AK{sub ye} and AK{sub ec} and the GMP complex of guanylate kinase the latter one leading to the definition of the monophosphate site, the problem of the location of the nucleotide sites can be considered to be solved in a way contradicting earlier work and denying the His residue homologous to H36 in porcine adenylate kinase a direct role in substrate binding.
- OSTI ID:
- 5557896
- Journal Information:
- Biochemistry; (United States), Vol. 30:17; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
NUCLEOTIDES
NUCLEAR MAGNETIC RESONANCE
PHOSPHOTRANSFERASES
CRYSTALLOGRAPHY
HISTIDINE
MOLECULAR STRUCTURE
OVERHAUSER EFFECT
PROTONS
SWINE
X-RAY DIFFRACTION
AMINO ACIDS
ANIMALS
AZOLES
BARYONS
CARBOXYLIC ACIDS
COHERENT SCATTERING
DIFFRACTION
DOMESTIC ANIMALS
ELEMENTARY PARTICLES
ENZYMES
FERMIONS
HADRONS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
IMIDAZOLES
MAGNETIC RESONANCE
MAMMALS
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
RESONANCE
SCATTERING
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques