FT-Infrared spectroscopic studies of the iron ligand CO stretch mode of iNOS oxygenase domain: effect of arginine and tetrahydrobiopterin.
The iron ligand CO stretch vibration mode of the inducible nitric oxide synthase oxygenase domain (iNOSox) has been studied from 20 to 298 K. iNOSox in the absence of arginine reveals a temperature-dependent equilibrium of two major conformational substates with CO stretch bands centered at about 1945 and 1954 cm(-)(1). This behavior is not qualitatively changed when tetrahydrobiopterin (H(4)B) is bound. Arginine binding changes significantly the spectrum by formation of a sharp CO stretch mode band at about 1905 cm(-)(1) and indicates the formation of a hydrogen bond to the CO ligand. For temperatures lower than 250 K, the stretch vibration frequency decreases almost linearly with decreasing temperature and indicates that the coupling between the CO ligand and the arginine/protein in the active site via the hydrogen bond is very strong. Flashphotolysis of the CO ligand carried out at 25 K revealed the CO stretch mode of the photodissociated CO ligand trapped in the heme pocket. There is a negative linear relation between the stretch vibration frequencies of the photodissociated and the iron-bound CO indicating that the photodissociated ligand stays near the heme.
Jung
C
C
Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Strasse Berlin, Germany. cjung@mdc-berlin.de
Stuehr
D J
DJ
Ghosh
D K
DK
D000818:Animals; D001120:Arginine / Q000494:pharmacology*; D001708:Biopterins / Q000031:analogs & derivatives* / Q000494:pharmacology; D002248:Carbon Monoxide / Q000737:chemistry*; D006420:Hemeproteins / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008956:Models, Chemical; D019001:Nitric Oxide Synthase / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects; D017550:Spectroscopy, Fourier Transform Infrared
D006420:Hemeproteins; D011994:Recombinant Proteins; D001708:Biopterins; D002248:Carbon Monoxide; D001120:Arginine; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C003402:sapropterin