19F nuclear magnetic resonance investigation of stereoselective binding of isoflurane to bovine serum albumin.
Whether proteins or lipids are the primary target sites for general anesthetic action has engendered considerable debate. Recent in vivo studies have shown that the S(+) and R(-) enantiomers of isoflurane are not equipotent, implying involvement of proteins. Bovine serum albumin (BSA), a soluble protein devoid of lipid, contains specific binding sites for isoflurane and other anesthetics. We therefore conducted 19F nuclear magnetic resonance measurements to determine whether binding of isoflurane to BSA was stereoselective. Isoflurane chemical shifts were measured as a function of BSA concentration to determine the chemical shift differences between the free and bound isoflurane. KD was determined by measuring the 19F transverse relaxation times (T2) as a function of isoflurane concentration. The binding duration was determined by assessing increases in 1/T2 as a result of isoflurane exchanging between the free and bound states. The S(+) and R(-) enantiomers exhibited no stereoselectivity in chemical shifts and KD values (KD = 1.3 +/- 0.2 mM, mean +/- SE, for S(+), R(-), and the racemic mixture). Nonetheless, stereoselectivity was observed in dynamic binding parameters; the S(+) enantiomer bound with slower association and dissociation rates than the R(-).
Xu
Y
Y
Department of Anesthesiology and Critical Care Medicine, University of Pittsburgh, Pennsylvania 15261, USA. y_xu@poplar.anes.upmc.edu
Tang
P
P
Firestone
L
L
Zhang
T T
TT
D018685:Anesthetics, Inhalation / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D005461:Fluorine / Q000737:chemistry; D066298:In Vitro Techniques; D007530:Isoflurane / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D013237:Stereoisomerism
D018685:Anesthetics, Inhalation; D012710:Serum Albumin, Bovine; D005461:Fluorine; D007530:Isoflurane