Binding of intermediate, product, and substrate analogs to neuronal nitric oxide synthase: ferriheme is sensitive to ligand-specific effects in the L-arginine binding site

Biochemistry. 1996 Sep 10;35(36):11839-45. doi: 10.1021/bi953015w.

Abstract

The electron paramagnetic resonance spectra of purified neuronal nitric oxide synthase indicates that the binding of ligands to the arginine site perturbs the environment of the high-spin ferriheme in a highly ligand-specific manner. Four categories of high-spin complex can be distinguished; all are five-coordinate, and all retain the axial thiolate ligand, but they differ in their ligation geometries. These spectroscopic species reveal distinct local conformations which can be stabilized individually by the binding of L-arginine, N omega-hydroxy-L-arginine, N omega-methyl-L-arginine, and N omega-nitro-L-arginine. Other arginine analog inhibitors stabilize one or more of these states, revealing patterns based on the nature of substituents at the terminal amino group.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Arginine / analogs & derivatives
  • Arginine / metabolism*
  • Arginine / pharmacology
  • Binding Sites
  • Cell Line
  • Citrulline / pharmacology
  • Electron Spin Resonance Spectroscopy
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacology
  • Heme / chemistry
  • Heme / metabolism*
  • Kidney
  • Neurons / enzymology*
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Nitric Oxide Synthase / chemistry
  • Nitric Oxide Synthase / metabolism*
  • Nitroarginine
  • Ornithine / analogs & derivatives
  • Ornithine / metabolism
  • Rats
  • omega-N-Methylarginine

Substances

  • Enzyme Inhibitors
  • Nitroarginine
  • omega-N-Methylarginine
  • Citrulline
  • N(G)-iminoethylornithine
  • N(5)-hydroxy-L-arginine
  • Heme
  • Arginine
  • Ornithine
  • Nitric Oxide Synthase