Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[<sup>3</sup>H]trimethisoquin

Saitoh, Tsunao; Oswald, Robert E.; Wennogle, Lawrence P.; Changeux, Jean‐Pierre

doi:10.1016/0014-5793(80)80522-9

Cited by 115 publications

(56 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Neutralization experiments were done under in vitro conditions. AcChR, prepared from electric organs from Torpedo marmorata according to [9], was incubated overnight at 4°C in the presence of radioactive toxins (protein concentrations are indicated in table 2) with various amounts of antibody in 1.5 ml Ringer buffer. The mixtures were filtered through two Millipore filters (HAWP, 0.45 pm) which were washed with 30 ml of 4°C Ringer solution.…”

Section: Methodsmentioning

confidence: 99%

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

et al. 1986

View full text Add to dashboard Cite

We isolated a neurotoxin-specific monoclonal antibody (Mab) which is capable of recognizing and neutralizing all short-chain toxin variants that have been tested, including those with widely divergent sequences. The epitope incorporates the three invariant residues Lys-27, Trp-29 and Lys-47 which form part of the site by which the toxins bind to the nicotinic acetylcholine receptor. To our knowledge, this is the first Mab which possesses the universal capacity of neutralizing all natural variants of a toxic protein. Monoclonal antibody Toxin Acetylcholine receptor

show abstract

Section: Methodsmentioning

confidence: 99%

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

et al. 1986

View full text Add to dashboard Cite

show abstract

“…Large batches (200 nmol of a-toxin-binding sites) of purified [15] and alkali-treated [16] AChR-rich membrane fragments were labeled according to Giraudat et al [2] with [3H]CPZ [2-3 Ci/mmol (preparation 1) or 12-14 Ci/mmol (preparation 2)1.…”

Section: Covalent Labeling Of the Achr By [~H]cpzmentioning

confidence: 99%

The noncompetitive blocker [³H]chlorpromazine labels segment M2 but not segment M 1 of the nicotinic acetylcholine receptor α‐subunit

et al. 1989

Self Cite

View full text Add to dashboard Cite

The membrane bound acetylcholine receptor from Torpedo marmorata was photolabeled by the noncompetitive channel blocker [~H]chlorpromazine under equilibrium conditions in the presence of the agonist carbamoylcholine. The radioactivity incorporated into the AChR subunits was reduced by addition of phencyclidine, a specific ligand for the high-affinity site for noncompetitive blockers. The ~t-subunit was purified and digested with trypsin and/or CNBr and the resulting fragments fractionated by HPLC. Sequence analysis resulted in the identification of Ser-248 as a major residue labeled by pH]chlorpromazine in a phencyclidine-sensitive manner. This residue is located in the hydrophobic and putative transmembrane segment M2 of the ~t-subunit, a region homologous to that containing the chlorpromazine-labeled Ser-262 in the Nchain [1] and the Ser-254 and Leu-257 in the//-chain [2]. Extended sequence analysis of the hydrophobic segment M 1 further showed that no labeling.occurred in this region.

show abstract

“…Postsynaptic membranes were prepared according to [20,21] from Torpedo marmorata electric organ in the presence of pretense inhibitots. The postsynaptie membranes were washed, concentrated by centrifugation and were used immediately after preparation.…”

Section: Methodsmentioning

confidence: 99%

Interaction of modified neurotoxins fromNaja nigricollis with the nicotinic acetylcholine receptor fromTorpedo marmorata A Raman spectroscopy study

et al. 1991

View full text Add to dashboard Cite

Two derivatives of or-toxin from Naja nigricollis venom were used in order to study, by resonance Raman spectroscopy, its interaction with tile nicotinic acetylcholine (AcCho) receptor from membranes of Torpedo marmorata electrocytes. The two modified toxins carry either an NO, group bound to Tyr 25 or a nitrophenylthioether (NPS) bound to Trp -'9, The comparison of the spectra of the free and bound derivatized toxins indicates that the environment of Tyr 2s is not perturbed upon binding to the AcCho receptor, but the surroundings of NPS bound to Tfp 29 are changed. This result indicates that Tyr "-s is not involved in binding, while Trp ~ of the ~-toxin may be in contact with the AcCho receptor. Examination of the spectrum of the AcCho receptor membrane after binding of the NPS-Trp toxin discloses some modifications ol ~ the vibrations of the tryptophan and cysteine disulfide bridge of the receptor. These residues are possibly involved in toxin binding.

show abstract

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[³H]trimethisoquin

Cited by 115 publications

References 29 publications

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

The noncompetitive blocker [³H]chlorpromazine labels segment M2 but not segment M 1 of the nicotinic acetylcholine receptor α‐subunit

Interaction of modified neurotoxins fromNaja nigricollis with the nicotinic acetylcholine receptor fromTorpedo marmorata A Raman spectroscopy study

Contact Info

Product

Resources

About

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[3H]trimethisoquin

Cited by 115 publications

References 29 publications

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

A monoclonal antibody which recognized the functional site of snake neurotoxins and which neutralizes all short‐chain variants

The noncompetitive blocker [3H]chlorpromazine labels segment M2 but not segment M 1 of the nicotinic acetylcholine receptor α‐subunit

Interaction of modified neurotoxins fromNaja nigricollis with the nicotinic acetylcholine receptor fromTorpedo marmorata A Raman spectroscopy study

Contact Info

Product

Resources

About

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[³H]trimethisoquin

The noncompetitive blocker [³H]chlorpromazine labels segment M2 but not segment M 1 of the nicotinic acetylcholine receptor α‐subunit