Nuclear magnetic relaxation studies of the role of the metal ion in Mn2(+)-substituted aminoacylase I.
Substitution of the essential Zn2+ ions of porcine kidney aminoacylase I (EC 3.5.1.14) by Mn2+ did not markedly affect the kinetic properties of the enzyme. Using Mn2+ as a paramagnetic probe, we were able to study the conformations of bound ligands by measuring the enhancement of ligand proton relaxation in 1H NMR. In addition, the effects of inhibitors on the paramagnetic enhancement of water proton relaxation rates were examined. The results of both approaches, in agreement with kinetic evidence, suggest that the metal center of aminoacylase I is too distant from the ligand binding site to allow direct participation of the metal in substrate binding or catalysis. We, therefore, propose that the metal ion of aminoacylase I plays a purely structural role.
Heese
D
D
Institut für Physiologische Chemie, Philipps-Universität, Marburg, Federal Republic of Germany.
Berger
S
S
Röhm
K H
KH
D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000502:physiology; D000818:Animals; D001665:Binding Sites; D004795:Enzyme Stability; D006860:Hydrogen Bonding; D007672:Kidney Cortex / Q000201:enzymology; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese / Q000032:analysis* / Q000502:physiology; D008433:Mathematics; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013552:Swine; D015032:Zinc / Q000502:physiology
D008345:Manganese; D000581:Amidohydrolases; C019672:aminoacylase I; D015032:Zinc