No abstract

No abstract

No abstract

Two photochromic activators of the electrogenic membrane of the electroplax of Electrophorus electricus are described. monium)methyljazobenzene dibromide (Bis-Q), one of the most potent ever reported, is active at concentrations of less than 10-7 M. Its cis isomer, which is obtained from the trans by exposure to light of 330 nm, is practically devoid of activity. Photoregulation of the potential of the membrane takes place in the presence of Bis-Q, presumably because of the conversion of the active trans isomer to the inactive cis isomer in the single-cell electroplax system.The second activator, 3-(a-bromomethyl)-3'-[a-(trimethylammonium)methyllazobenzene bromide (QBr) can be covalently attached to the electroplax membrane after reduction of the membrane with dithiothreitol. Activation of the membrane is induced by the covalently linked reagent. Its cis isomer, obtained from the trans by exposure to light of 330 nm, is, like cis-Bis-Q, of very low activity. Both isomers of Bis-Q are equally active as inhibitors of acetylcholinesterase, 50% inhibition occurring at a concentration of 10-5 M. The possibility of using trans-Bis-Q and trans-QBr to characterize and isolate the receptor protein is discussed.Systems in which photoregulation could be studied at the molecular level were described in previous papers. In these systems, photochromic azo derivatives were used as effector molecules to regulate the activities of chymotrypsin (1) and acetylcholinesterase (2, 3) and to photoregulate the potential of the excitable membrane of the monocellular electroplax preparation (4). Photoregulation was achieved by exploiting differences between the biochemical activities of the cis and trans isomers of the photochromic compounds, the relative concentrations of which were influenced by the wavelength of light to which the solution was exposed [or light vs. darkness, in one case (3) ].Light-induced changes in potential of the electroplax membrane may be considered as a Mtodel for the process of vision, in which the cis to trans isomerization of retinal is the first step in the initiation of a neural impulse. In the latter case, however, as well as in the phytochrome system of plants (5), the photochromic substances are located intracellularly, making for a highly efficient process. It thus appeared of interest to prepare a light-sensitive ligand that would form a covalent bond with the receptor protein of the electroplax. A compound with the desired properties was prepared: 3-(a-bromomethyl)-3'-[a-(trimethylammonium)methyl]azobenzene (QBr). Also synthesized was the closely related 3,3'-bis[a-(trimethylammonium)methyl]azobenzene (Bis-Q). the trans isomer of which was found to be a potent receptor activator, one of the most potent thus far described.The high affinity and specificity of Bis-Q may make it a useful reagent for the characterization, isolation, and purification of the receptor protein. Some experiments with the two azo compounds are presented in this paper. METHODSPreparation of 3,3'-bis(a-bromomethyl)azobenz...

No abstract