Characterization of sites occupied by the anesthetic nitrous oxide within proteins by infrared spectroscopy.
We report here a comprehensive infrared spectroscopic study of the interactions between the anesthetic nitrous oxide (N2O) and six proteins: lysozyme, cytochrome c, myoglobin, hemoglobin, serum albumin, and cytochrome c oxidase. Sites occupied by N2O molecules within these proteins were characterized. Three types of hydrophobic sites were found within the proteins. One with nu 3 near 2225 cm-1 is likely to be near peptide bond carbonyls; one with nu 3 near 2219 cm-1 may be near a benzene-like structure such as the side chains of phenylalanine and tyrosine; and the other with nu 3 near 2215 cm-1 is likely to be in a nonpolar alkane-like environment provided by the side chains of Leu, Ile, and Val residues. The amount of N2O molecules bound to myoglobin increases as the pH decreases from 9.2 to 5.2. N2O-protein interactions produced no detectable changes in the ligand-binding pockets of myoglobin, hemoglobin, and cytochrome c oxidase. N2O-induced secondary structure changes were detected only in the fully reduced cytochrome c oxidase, not in the fully oxidized oxidase and the other five proteins. N2O-induced conformational changes in the alpha beta-interface of hemoglobin and the h2 and h3 alpha-helices of human serum albumin were detected by monitoring the S-H stretch vibrations of cysteine residues. These findings provide direct evidence that anesthetic N2O interacts with proteins and occupies sites in the interior of the proteins.
Dong
A
A
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins 80523.
Huang
P
P
Zhao
X J
XJ
Sampath
V
V
Caughey
W S
WS
D000328:Adult; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009609:Nitrous Oxide / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012492:Salts; D013055:Spectrophotometry, Infrared
D011506:Proteins; D012492:Salts; D009609:Nitrous Oxide