Release of proteins from the inner surface of squid axon membrane labeled with tritiated N-ethylmaleimide.

Inoue, Ituro; Pant, Harish C.; Tasaki, Ichiji; Gainer, Harold

doi:10.1085/jgp.68.4.385

Cited by 29 publications

(17 citation statements)

References 15 publications

(18 reference statements)

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“…The Cl -effects are explained in terms of breakage of salt linkages among charged groups at the inner membrane layer by strong interactions between the charged groups and counterions having a large lyotropic number (cf. Tasaki, 1968 ;Inoue et al ., 1976). The rate of development of the adverse effects thus depends on the concentration of Cl-and on the duration of intracellular treatment with CI-.…”

Section: Methodsmentioning

confidence: 99%

Voltage-dependent chloride conductance of the squid axon membrane and its blockade by some disulfonic stilbene derivatives.

Inoue¹

1985

The Journal of General Physiology

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When giant axons of squid, Sepioteuthis, were bathed in a 100 mM Ca-salt solution containing tetrodotoxin (TTX) and internally perfused with a solution of 100 mM tetraethylammonium-salt (TEA-salt) or tetramethylammonium-salt (TMA-salt), the membrane potential was found to become sensitive to anions, especially Cl-. Membrane currents recorded from those axons showed practically no time-dependent properties, but they had a strong voltage-dependent characteristic, i.e., outward rectification. Cl- had a strong effect upon the voltage-dependent membrane currents. The nonlinear property of the currents was almost completely suppressed by some disulfonic stilbene derivatives applied intracellularly, such as 4-acetoamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) and as 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), which are blockers of chloride transport. On the basis of these experimental results, it is concluded that a voltage-dependent chloride-permeable channel exists in the squid axon membrane. The chloride permeability (PCl) is a function of voltage, and its value at the resting membrane (Em = -60 mV) is calculated, using the Goldman-Hodgkin-Katz equation, to be 3.0 X 10(-7) cm/s.

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Section: Methodsmentioning

confidence: 99%

Voltage-dependent chloride conductance of the squid axon membrane and its blockade by some disulfonic stilbene derivatives.

Inoue¹

1985

The Journal of General Physiology

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“…The supernatant (S) (KSCN removed proteins) as well as the pellet (P) (insoluble after KSCN) fractions were analyzed by Western blotting for ICln as well as the integral membrane protein Na-K-ATPase. Image shown is representative of those of two different experiments ripheral proteins [11] and then the pellet and the supernatant from this extraction were analyzed for ICln as well as for Na-K-ATPase, which is a known integral membrane protein. Extraction with the KSCN strips ICln from the particulate fraction (Fig.…”

Section: Distribution Of Icln and Translocation With Volume Expansionmentioning

confidence: 99%

Hypotonicity stimulates translocation of ICln in neonatal rat cardiac myocytes

Musch

Davis-Amaral

Vandenburgh

et al. 1998

Pfl�gers Archiv European Journal of Physiology

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Cell volume expansion stimulates the efflux of solutes, including the amino acid taurine, to accomplish a regulatory volume decrease (RVD). One protein that may play a role in taurine efflux is the cytosolic protein ICln. In rat neonatal cardiac myocytes under isotonic conditions, ICln is found predominantly (greater than 90%) in the cytosol. However, after cell volume expansion by exposure to hypotonic medium, ICln rapidly translocates to the particulate fraction (the Triton X-114-insoluble fraction). After 2 min in hypotonic medium the percentage of ICln in the particulate fraction increases to 30%, 46% at 5 min, 40% at 10 min, and 25% at 30 min. The time course of this response is similar to that of hypotonicity-stimulated taurine efflux. Hypotonicity-stimulated taurine efflux as well as ICln translocation parallel the reduction in medium osmolarity. As osmolarity decreases, taurine efflux and ICln movement increase. The movement of ICln from the particulate back to the cytosolic fraction is accelerated when volume-expanded cells are returned to isotonic medium. When ICln is analyzed under non-denaturing conditions, a dimer is detected in the particulate fraction of volume-expanded cells, along with the monomer. This dimer is not detected in the cytosol. Treatment of the particulate fraction from volume-expanded cells with the lyotropic agent KSCN caused release of ICln but not Na-K-ATPase into the soluble fraction, indicating that translocated ICln associates with membranes in the particulate fraction rather than inserting into them.

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“…Electron microscopically, Metuzals and Tasaki (1978) showed that a three-dimensional network of interwoven filaments consisting partly of an actin-like protein is firmly attached to the axolemma and might have a role in some aspect of excitability. Upon analyzing axonal proteins in perfusate with SDS-polyacrylamide gel electrophoresis, Gainer et al (1974), Inoue et aL (1976), and Takenaka et al (1976) concluded that a 12,000-dalton protein was released after repetitive electrical stimulation of the axon or by potassium depolarization. However, quite recently, Pant et al (1978) concluded that an appreciable amount of a 45,000-dalton protein, in addition to the 12,000 daltons, presented in the perfusate from the stimulated axon or the axon exhibiting long-lasting action potential, while a 68,000-dalton protein dominated in the perfusate from the depolarized axon.…”

mentioning

confidence: 99%

Microtubules inside the plasma membrane of squid giant axons and their possible physiological function

Matsumoto¹,

Sakai²

1979

J. Membrain Biol.

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The effects of application of the microtubule-disassembling reagents to squid giant axons upon resting potential, the height of the propagated action potential, and the threshold to evoke action potential were studied using colchicine, podophyllotoxin, vinblastine, griseofulvin, sulfhydryl reagents including NEM, diamide, DTNB and PCMB, and Ca2+ ions. At the same time, the effects of concentrations of K halides and K glutamate on the above physiological properties were studied in comparison with in vitro characteristics of microtubule assembly from purified axoplasmic tubulin. It was found that there was good correlation between conditions supporting maintenance of membrane excitability and microtubule assembly. The experiments suggest that associated with the internal surface of the plasma membrane there are microtubules which regulate in part both the resting and action potentials.

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Release of proteins from the inner surface of squid axon membrane labeled with tritiated N-ethylmaleimide.

Cited by 29 publications

References 15 publications

Voltage-dependent chloride conductance of the squid axon membrane and its blockade by some disulfonic stilbene derivatives.

Voltage-dependent chloride conductance of the squid axon membrane and its blockade by some disulfonic stilbene derivatives.

Hypotonicity stimulates translocation of ICln in neonatal rat cardiac myocytes

Microtubules inside the plasma membrane of squid giant axons and their possible physiological function

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