Inhalational anesthetic-binding proteins in rat neuronal membranes.
Molecular targets of inhaled anesthetics must be represented in the group that specifically bind these drugs, but the paucity of direct binding data has limited the number of candidates for further evaluation. To find candidate targets, we used a combination of photolabeling, two-dimensional gel electrophoresis, and mass spectrometry to identify halothane-binding targets in rat neuronal membranes. Of the 265 spots detected on the two-dimensional gels, 90 were labeled by [(14)C]halothane, and 34 were identified. Mitochondrial proteins, especially respiratory complex and voltage-dependent anion channels, dominated the labeled group, and there were several examples of subunit- and state-dependent binding. A significant correlation was found between internal protein cavities and binding in a group of proteins with high resolution structures. Therefore, in addition to identifying novel neuronal targets, these data suggest a general molecular feature, the buried cavity, as a dominant attribute of volatile anesthetic-binding sites found in a limited number of neuronal membrane proteins.
Department of Anesthesia, University of Pennsylvania Health System, 305 John Morgan, 3620 Hamilton Walk, Philadelphia, PA 19104, USA.
Jin
J
Xi
Renyu
R
Liu
G Reid
GR
Asbury
Maryellen F
MF
Eckenhoff
Roderic G
RG
Eckenhoff
D000777:Anesthetics / Q000737:chemistry*; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins; D002462:Cell Membrane / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006221:Halothane / Q000737:chemistry; D008297:Male; D013058:Mass Spectrometry; D008928:Mitochondria / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000777:Anesthetics; D018685:Anesthetics, Inhalation; D002352:Carrier Proteins; D006221:Halothane