Entry of diphtheria toxin into cells: possible existence of cellular factor(s) for entry of diphtheria toxin into cells was studied in somatic cell hybrids and hybrid toxins.

Kaneda, Yasufumi; Uchida, Tsuyoshi; Mekada, Eisuke; Nakanishi, Masami; Okada, Yoshio

doi:10.1083/jcb.98.2.466

Cited by 22 publications

(13 citation statements)

References 32 publications

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“…HeLa cells are ,~100 times and mouse L cells ~105 times less sensitive than Vero cells (28). Ehrlich ascites tumor cells are more resistant than L cells (15). Fig.…”

Section: Cell Membranes Contain An Inhibitor Of Dt Cytotoxicitymentioning

confidence: 99%

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Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane.

Mekada

Okada

Uchida

1988

The Journal of Cell Biology

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Abstract. Two substances possessing the ability to bind to diphtheria toxin (DT) were found to be present in a membrane fraction from DT-sensitive Vero cells. One of these substances was found on the basis of its ability to bind DT and inhibit its cytotoxic effect. This inhibitory substance competitively inhibited the binding of DT to Veto cells. However this inhibitor could not bind to CRM197, the product of a missense mutation in the DT gene, and did not inhibit the binding of CRM197 to Vero cells. Moreover, similar levels of the inhibitory activity were observed in membrane fractions from DT-insensitive mouse cells, suggesting the inhibitor is not the DT receptor which is specifically present in DT-sensitive cells. The second DT-binding substance was found in the same Vero cell membrane preparation by assaying the binding of ~25I-labeled CRM197. Such DT-binding activity could not be observed in membrane preparation from mouse L cells. From competition studies using labeled DT and CRM proteins, we conclude that this binding activity is due to the surface receptor for DT. Treatment of these substances with several enzymes revealed that the inhibitor was sensitive to certain RNases but resistant to proteases, whereas the DT receptor was resistant to RNase but sensitive to proteases. The receptor was solubilized and partially purified by chromatography on CM-Sepharose column. Immunoprecipitation and Western blotting analysis of the partially purified receptor revealed that a 14.5-kD protein is the DT receptor, or at least a component of it.D PHTHERIA toxin (DT) ~ is a cytotoxic protein which inhibits cellular protein synthesis (4, 40) in eukaryotes by catalyzing the ADP-ribosylation of EF-2, which results in its inactivation (9, 12). The first step of intoxication by DT is binding of the toxin to a susceptible cell. A specific receptor for DT is believed to be involved in this step (13,43). Cells from a number of mammals including humans and monkeys are sensitive to DT, but mouse and rat cells are insensitive (29). Several lines of evidence show that the difference in sensitivity to DT between species is primarily determined by the presence or absence of a cell surface receptor (18,28,31,48). However, this receptor has not been isolated. The toxins bound to cell surface are then internalized by endocytosis (33), and the toxins, or at least their A fragments, enter the cytoplasm to exert its effects. Like in the case of Semliki Forest virus (SFV) (11), intravesicular low pH is required for the penetration of the toxin into cytoplasm (6,17,20,27,36).Some information has been obtained on the biochemical properties of DT receptor. The treatment of DT-sensitive cells with some proteases or phospholipase C reduces the sensitivity to DT (32), whereas treatment with neuraminidase increases the sensitivity (26). Although these studies 1. Abbreviations used in this paper: CRMs, cross reacting materials related to diphtheria toxin; DT, diphtheria toxin.give a clue to the chemical nature of DT receptor, the possibility that ...

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“…HeLa cells are ,~100 times and mouse L cells ~105 times less sensitive than Vero cells (28). Ehrlich ascites tumor cells are more resistant than L cells (15). Fig.…”

Section: Cell Membranes Contain An Inhibitor Of Dt Cytotoxicitymentioning

confidence: 99%

“…The nicked form of CRM197 was prepared by treatment with trypsin (7). Fragment A of diphtheria toxin was purified from the culture fluid of the C7 (15)(16)(17)(18)(19)(20)(21)(22) strain (45).…”

Section: Dt and Related Proteinsmentioning

confidence: 99%

Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane.

Mekada

Okada

Uchida

1988

The Journal of Cell Biology

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“…In the present study, we used a viral protein-liposome-mediated ODN transfer technique that made use of the liposome and the protein coat of the inactivated Sendai virus (hemagglutinating virus of Japan [HVJ]). 12,13 HVJ contains 2 envelope proteins, hemagglutinin-neuraminidase and fusion proteins, which mediate cell attachment and membrane fusion sequentially. 14,15 In the present study, we reasoned that if tissue ACE contributes to neointimal formation, we would observe a reduction of lesion formation by antisense ODNs in the absence of any effects on systemic blood pressure or the circulating renin-angiotensin system.…”

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confidence: 99%

Antisense Oligodeoxynucleotide Inhibition of Vascular Angiotensin-Converting Enzyme Expression Attenuates Neointimal Formation

Morishita

Gibbons

Tomita

et al. 2000

ATVB

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Abstract-It has been proposed that vascular angiotensin-converting enzyme (ACE) plays an important role in regulating vascular growth. Indeed, ACE inhibitors have been reported to prevent neointimal formation after vascular injury in a rat carotid artery model. However, classic pharmacological experiments cannot exclude the potential contributions of hemodynamics and the circulating renin-angiotensin system (RAS). In this study, we used antisense oligodeoxynucleotide (ODN) to obtain local blockade of vascular ACE expression without effects on systemic hemodynamics and circulating RAS. To increase the effectiveness of antisense action, we modified the hemagglutinating virus of Japan-liposome ODN delivery method by cotransfection with nuclear protein (high mobility group 1 [HMG-1]) and RNase H. In vitro experiments showed the enhanced efficacy of antisense ODN by cotransfection of HMG-1 and RNase H compared with ODN alone. In vivo transfection of antisense ACE ODNs into intact uninjured rat carotid artery resulted in a significant reduction of vascular ACE activity, and cotransfection of HMG-1 and RNase H showed further reduction. We examined the effects of local blockade of vascular ACE expression on neointimal formation after vascular injury. Transfection of antisense ACE ODNs resulted in the attenuation of neointimal formation, whereas sense and scrambled ODNs did not. Blood pressure, heart rate, and serum ACE activity were not affected by antisense treatment. Key Words: gene therapy Ⅲ restenosis Ⅲ renin-angiotensin system Ⅲ antisense oligonucleotides Ⅲ hemagglutinating virus of Japan O ne of the current controversies in cardiovascular research is the existence of a functional tissue angiotensin system 1-4 in the blood vessel. The presence of a vascular angiotensin system has been reported previously. [5][6][7] Furthermore, we have described that vascular angiotensin-converting enzyme (ACE) expression is induced in the intimal smooth muscle cells of rat carotid artery and abdominal aorta in response to vascular injury. 8,9 This injury model has provided us with the opportunity to examine the functional role of local angiotensin production. Using a dose range of an ACE inhibitor, we demonstrated that the magnitude of neointimal lesion reduction correlated closely with the degree of vascular ACE inhibition. 10 We have interpreted these data as supportive evidence for a functional role of tissue angiotensin. However, the blockade of tissue angiotensin required an extremely high dosage of ACE inhibitors, which was invariably accompanied by the suppression of circulating renin angiotensin and the reduction of systemic blood pressure. Therefore, it is still unclear whether the local blockade of vascular angiotensin can indeed prevent neointimal formation without producing a systemic blockade of circulating renin angiotensin and/or without affecting hemodynamics. Furthermore, ACE inhibition also results in bradykinin accumulation and nitric oxide induction, which may contribute to the inhibition of neointimal hyperpla...

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“…Cholera toxin B fragment and wheat germ agglutinin are highly sensitive retrograde tracers [184][185][186]. Neurotoxins, including diphtheria toxin, botulinum toxin, ricin, and saporin, have been conjugated with these retrograde tracers to kill connected neurons [187][188][189][190]. For example, respiratory motor neurons were with intrapleural injections of cholera toxin Bconjugated saporin [190].…”

Section: Cell-specific Neurotoxinsmentioning

confidence: 99%

Selective Manipulation of Neural Circuits

Park

Carmel

2016

Neurotherapeutics

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Unraveling the complex network of neural circuits that form the nervous system demands tools that can manipulate specific circuits. The recent evolution of genetic tools to target neural circuits allows an unprecedented precision in elucidating their function. Here we describe two general approaches for achieving circuit specificity. The first uses the genetic identity of a cell, such as a transcription factor unique to a circuit, to drive expression of a molecule that can manipulate cell function. The second uses the spatial connectivity of a circuit to achieve specificity: one genetic element is introduced at the origin of a circuit and the other at its termination. When the two genetic elements combine within a neuron, they can alter its function. These two general approaches can be combined to allow manipulation of neurons with a specific genetic identity by introducing a regulatory gene into the origin or termination of the circuit. We consider the advantages and disadvantages of both these general approaches with regard to specificity and efficacy of the manipulations. We also review the genetic techniques that allow gain-and loss-of-function within specific neural circuits. These approaches introduce light-sensitive channels (optogenetic) or drug sensitive channels (chemogenetic) into neurons that form specific circuits. We compare these tools with others developed for circuit-specific manipulation and describe the advantages of each. Finally, we discuss how these tools might be applied for identification of the neural circuits that mediate behavior and for repair of neural connections.

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Entry of diphtheria toxin into cells: possible existence of cellular factor(s) for entry of diphtheria toxin into cells was studied in somatic cell hybrids and hybrid toxins.

Cited by 22 publications

References 32 publications

Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane.

Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane.

Antisense Oligodeoxynucleotide Inhibition of Vascular Angiotensin-Converting Enzyme Expression Attenuates Neointimal Formation

Selective Manipulation of Neural Circuits

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