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

H A Oktem; J Moitra; S Benyhe; G Toth; A Lajtha; A Borsodi

doi:10.1016/0024-3205(91)90214-v

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

Life Sci. 1991;48(18):1763-8. doi: 10.1016/0024-3205(91)90214-v.

Authors

H A Oktem¹, J Moitra, S Benyhe, G Toth, A Lajtha, A Borsodi

Affiliation

¹ Institute of Biochemistry, Hungarian Academy of Sciences, Szeged.

PMID: 1850495
DOI: 10.1016/0024-3205(91)90214-v

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Chloromethyl Ketones / metabolism*
Animals
Brain / metabolism*
Cell Membrane / metabolism
Centrifugation, Density Gradient
Chromatography, Gel
Electrophoresis, Polyacrylamide Gel
Molecular Weight
Rats
Rats, Inbred Strains
Receptors, Opioid / isolation & purification
Receptors, Opioid / metabolism*
Receptors, Opioid, mu
Tritium

Substances

Amino Acid Chloromethyl Ketones
Receptors, Opioid
Receptors, Opioid, mu
Tritium
tyrosyl-alanyl-glycyl-N(alpha)-methylphenylalanine chloromethyl ketone