31P and 1H NMR studies of the structure of enzyme-bound substrate complexes of lobster muscle arginine kinase: relaxation measurements with Mn(II) and Co(II)

• Published in: Biochemistry (Easton) 1989-11, Vol.28 (24), p.9343-9350
• Main Authors: JARORI, G. K, RAY, B. D, NAGESWARA RAO, B. D
• Format: Article
• Language: English
• Subjects: Adenosine Diphosphate - metabolism; Adenosine Triphosphate - metabolism; Animals; Arginine Kinase - metabolism; Biological and medical sciences; Chemical Phenomena; Chemistry, Physical; Cobalt - metabolism; Electron Spin Resonance Spectroscopy; Fundamental and applied biological sciences. Psychology; Hydrogen; Magnetic Resonance Spectroscopy; Manganese - metabolism; Molecular biophysics; Muscles - enzymology; Nephropidae; Phosphorus; Phosphotransferases - metabolism; Rabbits; Spectroscopy : techniques and spectras; Temperature; Thermodynamics
• ISSN: 0006-2960; 1520-4995

The paramagnetic effects of Mn(II) and Co(II) on the spin-lattice relaxation rates of 31P nuclei of ATP and ADP and of Mn(II) on the spin-lattice relaxation rate of the delta protons of arginine bound to arginine kinase from lobster tail muscle have been measured. Temperature variation of 31P relaxation rates in E.MnADP and E.MnATP yields activation energies (delta E) in the range 6-10 kcal/mol. Thus, the 31P relaxation rates in these complexes are exchange limited and cannot provide structural information. However, the relaxation rates in E.CoADP and E.CoATP exhibit frequency dependence and delta E values in the range 1-2 kcal/mol; i.e., these rates depend upon 31P-Co(II) distances. These distances were calculated to be in the range 3.2-4.5 A, appropriate for direct coordination between Co(II) and the phosphoryl groups. The paramagnetic effect of Mn(II) on the 1H spin-lattice relaxation rate of the delta protons of arginine in the E.MnADP.Arg complex was also measured at three frequencies (viz., 200, 300, and 470 MHz). These 1H experiments were performed in the presence of sufficient excess of arginine to be observable over the protein background but with MnADP exclusively in the enzyme-bound form so that the enhancement in the relaxation rates of the delta protons of arginine arises entirely from the enzyme-bound complex. Both the observed frequency dependence of these rates and the delta E less than or equal to 1.0 +/- 0.3 kcal/mol indicate that this rate depends on the 1H-Mn(II) distances.