Identification in the mu-opioid receptor of cysteine residues responsible for inactivation of ligand binding by thiol alkylating and reducing agents

FEBS Letters
G GaibeletL J Emorine

Abstract

Inactivation by thiol reducing and alkylating agents of ligand binding to the human mu-opioid receptor was examined. Dithiothreitol reduced the number of [3H]diprenorphine binding sites. Replacement by seryl residues of either C142 or C219 in extracellular loops 1 and 2 of the mu receptor resulted in a complete loss of opioid binding. A disulfide bound linking C142 to C219 may thus be essential to maintain a functional conformation of the receptor. We also demonstrated that inactivation of ligand binding upon alkylation by N-ethylmaleimide occurred at two sites. Alteration of the more sensitive (IC50 = 20 microM) did not modify antagonists binding but decreased agonist affinity almost 10-fold. Modification of the less reactive site (IC50 = 2 mM) decreased the number of both agonist and antagonist binding sites. The alkylation site of higher sensitivity to N-ethylmaleimide was shown by mutagenesis experiments to be constituted of both C81 and C332 in transmembrane domains 1 and 7 of the mu-opioid receptor.

References

Jun 9, 1977·Nature·J A LordH W Kosterlitz
Jun 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·E J Simon, J Groth
Dec 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·B L KiefferC G Hirth
Dec 28, 1992·Science·C J EvansR H Edwards
Jan 1, 1991·Life Sciences·S R Childers
Nov 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·S S KarnikH G Khorana
Jul 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·E J SimonI Edelman
Nov 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·J B WangG R Uhl
Nov 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·F MengH Akil
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·K YasudaG I Bell
Aug 9, 1996·The Journal of Biological Chemistry·D E KeithM von Zastrow

❮ Previous
Next ❯

Citations

Mar 26, 1999·European Journal of Immunology·G MacéG Dietrich
Aug 7, 2002·European Journal of Pharmacology·Faika MseehMargarita I Dubocovich
Mar 8, 2000·Peptides·L M Harrison, D K Grandy
Mar 13, 2003·Hypertension Research : Official Journal of the Japanese Society of Hypertension·Kenji KuwasakoTanenao Eto
Apr 11, 2015·ACS Chemical Biology·Dietmar Weichert, Peter Gmeiner
Feb 27, 2004·Protein Engineering, Design & Selection : PEDS·David OttAndreas Plückthun
Jul 9, 2003·Journal of Peptide Science : an Official Publication of the European Peptide Society·Kálmán Medzihradszky
Apr 24, 1999·European Journal of Biochemistry·G GaibeletL J Emorine

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.