Interaction of tritium-labeled H2DIDS (4,4′-diisothiocyano-1,2,diphenyl ethane-2,2′disulfonic acid) with the ehrlich mouse ascites tumor cell

Levinson, Charles; Corcoran, Rebecca J.; Edwards, Ellen H.

doi:10.1007/bf01869295

Cited by 22 publications

(6 citation statements)

References 27 publications

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“…Group B streptococci could also be labeled with the surface label [3H]DIDS as described by Levinson et al (11). The binding of these 3Hlabeled streptococci to the infected MDCK cells was proportional to the number of streptococci added to about 5 mg of cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…[14C]mannose (300 uCi/iLmol) was from Research Products International Co. 4,4'-[3H]diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid ([3H]DIDS) for labeling the streptococci was kindly supplied by Charles Lev-inson, Department of Physiology of this institution, as was the procedure for labeling (11). Tumncamycin was a generous gift from Robert Hamill, Eli Lilly & Co., and streptovirudin was kindly supplied by K. Eckardt, Zentralinstitut fur Mikrobiolgie und Experimentalle Therapie, Jena, East Germany.…”

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confidence: 99%

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Adherence of bacteria to mammalian cells: inhibition by tunicamycin and streptovirudin

Pan¹,

Schmitt²,

Sanford³

et al. 1979

J Bacteriol

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Group B streptococci were labeled either by growing the cells in ['4C]fructose or by using the surface label 4,4'-[3H]diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid, which reacts with amino groups. A quantitative assay was developed by using these labeled bacteria to study the adherence of streptococci to canine kidney epithelial celLs. The bacteria adhered to kidney celLs that had been infected with influenza A virus, but did not adhere to uninfected cells. The binding of 3Hlabeled group B streptococci was proportional to the number of bacteria added and showed saturation kinetics. The binding was blocked by the addition of unlabeled group B streptococci but was not affected by addition of streptococci from other groups. It was also blocked by mixing the 3H-labeled streptococci with influenza A virus before adding the bacteria to the kidney cells. When the kidney cells were infected with influenza virus in the presence of either tunicamycin or streptovirudin, these antibiotics inhibited the appearance of viral hemagglutinin in the kidney cells and also prevented the release of mature virus. In these experiments, the adherence of 'H-labeled streptococci was also inhibited. Tunicamycin was shown to block the incorporation of [14C]mannose into lipid-linked oligosaccharides and glycoprotein in both normal and virus-infected kidney cells. These data give strong support to the notion that adherence of streptococci to mammali cells involves recognition ofviral hemagglutinin, a glycoprotein whose synthesis is blocked by certain antibiotics.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Adherence of bacteria to mammalian cells: inhibition by tunicamycin and streptovirudin

Pan¹,

Schmitt²,

Sanford³

et al. 1979

J Bacteriol

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“…1), indicating that DIDS acts on the outside of the cell membrane, as it does in the human red blood cell (see Deuticke, 1977). The permeability to 4,4'-diisothiocyano-l,2-diphenylethane-2,2'-disulfonic acid (H2DIDS) has been shown to be low in the Ehrlich cell (Levinson et al, 1979). The irreversible binding of DIDS is low within the first 2 min.…”

Section: Discussionmentioning

confidence: 97%

Identification of the anion exchange protein of ehrlich cells: A kinetic analysis of the inhibitory effects of 4,4′-diisothiocyano-2,2′-stilbene-disulfonic acid (DIDS) and labeling of membrane proteins with3H-DIDS

1986

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In Ehrlich ascites tumor cells 4,4'-diisothiocyano-2,2'-stilbene-disulfonic acid (DIDS) inhibits the chloride exchange both reversibly and irreversibly. The reversible inhibition is practically instantaneous and of a competitive nature with Ki about 2 microM at zero chloride concentration. This is succeeded by a slow irreversible binding of DIDS to the transporter, with a chloride dependence suggesting binding to the same site as for reversible DIDS binding/inhibition. To identify the membrane protein involved in anion exchange, cells were labeled with 3H-DIDS. Incubation of cells for 10 min with 25 microM DIDS at pH 8.2 leads to more than 95% inhibition of the DIDS-sensitive chloride exchange flux when the chloride concentration is low (15 mM). This condition was used for the 3H-DIDS-labeling experiments. After incubation the cells were disrupted, the membranes isolated and solubilized, and the proteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The distribution of the 3H-activity in the gel showed only one major peak, which could be related to protein with a mol wt of about 30,000 Daltons. The number of transport sites was estimated at about 400,000 per cell, and from the DIDS-sensitive chloride flux under steady-state conditions we calculate a turnover number of 340 ions per sec per site.

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“…H2DIDS and [3H]H2DIDS were made in our laboratory (Jennings et al, 1984) according to methods adapted from Lepke et al (1976) and Levinson et al (1979). BS3 was made according to the method of Staros (1982b) or was purchased from Pierce.…”

Section: Methodsmentioning

confidence: 99%

N-hydroxysulfosuccinimido active esters and the L-(+)-lactate transport protein in rabbit erythrocytes

Donovan¹,

Jennings²

1986

Biochemistry

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Esters of N-hydroxysulfosuccinimide strongly inhibit L-(+)-lactate transport in rabbit erythrocytes, probably by acylating amino groups on the transport protein. Lactate transport studies using bis(sulfosuccinimido) suberate (BS3), bis(sulfosuccinimido) adipate (BS2A), bis(sulfosuccinimido) dithiobis(propionate), and a variety of monocarboxylate esters suggest that an exofacial amino group of the lactate transport protein is essential for lactate transport. Also, reductive methylation studies show that even when positive charge is preserved in modified amino groups, the transport is strongly inhibited. At pH less than 6, band 3 mediated inorganic anion transport is enhanced in BS3-treated cells, while at pH greater than 6, it is inhibited. BS3-induced inhibition of L-(+)-lactate transport does not have this pH dependence. BS3 reduces the labeling of a 40-50-kDa membrane polypeptide (band R) by tritiated 4,4'-diisothiocyanato-2,2-dihydrostilbenedisulfonate ([3H]H2DIDS) and by tritiated bis(sulfosuccinimido) adipate ([3H]BS2A). Tritiated sulfosuccinimido acetate (S2[3H]acetate) also labels band R, over a range of concentrations where lactate transport is inhibited in a dose-dependent manner by S2 acetate. BS3 is a known impermeant protein cross-linker. S2 acetate permeates rabbit red cell membranes by an H2DIDS-inhibitable mechanism. BS3 cross-links the proteolytic fragments of rabbit band 3 produced by extracellular chymotrypsin. These labeling experiments support an association between band R and specific monocarboxylate transport.

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Interaction of tritium-labeled H2DIDS (4,4′-diisothiocyano-1,2,diphenyl ethane-2,2′disulfonic acid) with the ehrlich mouse ascites tumor cell

Cited by 22 publications

References 27 publications

Adherence of bacteria to mammalian cells: inhibition by tunicamycin and streptovirudin

Adherence of bacteria to mammalian cells: inhibition by tunicamycin and streptovirudin

Identification of the anion exchange protein of ehrlich cells: A kinetic analysis of the inhibitory effects of 4,4′-diisothiocyano-2,2′-stilbene-disulfonic acid (DIDS) and labeling of membrane proteins with3H-DIDS

N-hydroxysulfosuccinimido active esters and the L-(+)-lactate transport protein in rabbit erythrocytes

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