Opioid receptor labeling with the chloromethyl ketone derivative of [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol (DAMGO) II: Covalent labeling of mu opioid binding site by 3H-Tyr-D-Ala-Gly-(Me)Phe chloromethyl ketone.
Opioid receptors of rat brain membranes were prelabeled with 3H-Tyr-D-Ala2-(Phe4)-Gly-CH2Cl, a chloromethyl ketone derivative of enkephalin, and solubilized in 1% digitonin. Hydrodynamic parameters of the receptor detergent complex derived from gel filtration and sucrose density gradient ultracentrifugation were found to be 51 A and 8.7 S, respectively, and the size was estimated to be about 200 kDa. Sodium dodecyl sulfate gel electrophoresis followed by fluorography revealed specific alkylation of a major protein at 58 kDa.
Oktem
H A
HA
Institute of Biochemistry, Hungarian Academy of Sciences, Szeged.
Moitra
J
J
Benyhe
S
S
Toth
G
G
Lajtha
A
A
Borsodi
A
A
D000590:Amino Acid Chloromethyl Ketones / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011957:Receptors, Opioid / Q000302:isolation & purification / Q000378:metabolism*; D017450:Receptors, Opioid, mu; D014316:Tritium
D000590:Amino Acid Chloromethyl Ketones; D011957:Receptors, Opioid; D017450:Receptors, Opioid, mu; D014316:Tritium; C052900:tyrosyl-alanyl-glycyl-N(alpha)-methylphenylalanine chloromethyl ketone