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NMR studies of the AMP-binding site and mechanism of adenylate kinase

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00380a024· OSTI ID:6410051
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NMR has previously been used to determine the conformation of enzyme-bound MgATP and to locate the MgATP-binding site on adenylate kinase. To determine the conformation and location of the other substrate, AMP, distances have been measured from Cr/sup 3 +/AMPPCP, a linear competitive inhibitor with respect to MgATP, to six protons and to the phosphorus atom of AMP on adenylate kinase, with the paramagnetic probe-T/sub 1/ method. Time-dependent nuclear Overhauser effects (NOEs) have been used to measure five interproton distances on enzyme-bound AMP. These distances were used to determine the conformation of bound AMP in addition to its position with respect to metal-ATP. Ten intermolecular NOEs, from protons of the enzyme to those of AMP, were detected, indicating the proximity of at least three hydrophobic amino acids to bound AMP. These constraints, together with the conformation of AMP and the intersubstrate distances, were used to position AMP into the X-ray structure of adenylate kinase. The AMP binding site is found to be near Leu-116, Arg-171, Val-173, Val-182, and Leu-190; all of these residues have been found to be invariant in muscle-type rabbit, calf, human, porcine.

Research Organization:
Johns Hopkins Univ. School of Medicine, Baltimore, MD
OSTI ID:
6410051
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:6; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550601 -- Medicine-- Unsealed Radionuclides in Diagnostics
550602* -- Medicine-- External Radiation in Diagnostics-- (1980-)
62 RADIOLOGY AND NUCLEAR MEDICINE
AMP
BIOCHEMICAL REACTION KINETICS
COHERENT SCATTERING
CONFIGURATION INTERACTION
DIFFRACTION
ENZYMES
ISOTOPES
KINETICS
LIGHT NUCLEI
MAGNETIC RESONANCE
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
OVERHAUSER EFFECT
PHOSPHORUS 31
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
REACTION KINETICS
RESONANCE
SCATTERING
SPECTRA
STABLE ISOTOPES
TRANSFERASES
X-RAY DIFFRACTION