Bacitracin inhibits the reductive activity of protein disulfide isomerase by disulfide bond formation with free cysteines in the substrate‐binding domain

Dickerhof, Nina; Kleffmann, Torsten; Jack, Ralph W.; McCormick, Sally P.A.

doi:10.1111/j.1742-4658.2011.08119.x

Cited by 62 publications

(50 citation statements)

References 29 publications

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“…These observations provide proof of principle for targeting extracellular PDI for inhibition of thrombus formation. Other agents that inhibit PDI function, such as juniferdin (14) or bacitracin (45), also inhibit thrombus formation in vivo (data not shown and ref. 7).…”

Section: Figurementioning

confidence: 99%

Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents

Jasuja

Passam²,

Kennedy³

et al. 2012

J. Clin. Invest.

259

338

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Thrombosis, or blood clot formation, and its sequelae remain a leading cause of morbidity and mortality, and recurrent thrombosis is common despite current optimal therapy. Protein disulfide isomerase (PDI) is an oxidoreductase that has recently been shown to participate in thrombus formation. While currently available antithrombotic agents inhibit either platelet aggregation or fibrin generation, inhibition of secreted PDI blocks the earliest stages of thrombus formation, suppressing both pathways. Here, we explored extracellular PDI as an alternative target of antithrombotic therapy. A high-throughput screen identified quercetin-3-rutinoside as an inhibitor of PDI reductase activity in vitro. Inhibition of PDI was selective, as quercetin-3-rutinoside failed to inhibit the reductase activity of several other thiol isomerases found in the vasculature. Cellular assays showed that quercetin-3-rutinoside inhibited aggregation of human and mouse platelets and endothelial cell-mediated fibrin generation in human endothelial cells. Using intravital microscopy in mice, we demonstrated that quercetin-3-rutinoside blocks thrombus formation in vivo by inhibiting PDI. Infusion of recombinant PDI reversed the antithrombotic effect of quercetin-3-rutinoside. Thus, PDI is a viable target for small molecule inhibition of thrombus formation, and its inhibition may prove to be a useful adjunct in refractory thrombotic diseases that are not controlled with conventional antithrombotic agents.

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Section: Figurementioning

confidence: 99%

Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents

Jasuja

Passam²,

Kennedy³

et al. 2012

J. Clin. Invest.

259

338

View full text Add to dashboard Cite

show abstract

“…Several small molecules were previously reported as selective irreversible PDI inhibitors that suppressed apoptosis caused by misfolded proteins in a model of Huntington disease (4). A peptide antibiotic, bacitracin, interacts with and inhibits PDI through disulfide bond formation with activity in the high micromolar range (16). Although bacitracin is widely used as a PDI inhibitor in research, its clinical use is hampered by its nephrotoxicity and low membrane permeability (17)(18)(19).…”

mentioning

confidence: 99%

Discovery of an orally active small-molecule irreversible inhibitor of protein disulfide isomerase for ovarian cancer treatment

Butkevich

Yamada

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

188

212

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Protein disulfide isomerase (PDI), an endoplasmic reticulum chaperone protein, catalyzes disulfide bond breakage, formation, and rearrangement. The effect of PDI inhibition on ovarian cancer progression is not yet clear, and there is a need for potent, selective, and safe small-molecule inhibitors of PDI. Here, we report a class of propynoic acid carbamoyl methyl amides (PACMAs) that are active against a panel of human ovarian cancer cell lines. Using fluorescent derivatives, 2D gel electrophoresis, and MS, we established that PACMA 31, one of the most active analogs, acts as an irreversible small-molecule inhibitor of PDI, forming a covalent bond with the active site cysteines of PDI. We also showed that PDI activity is essential for the survival and proliferation of human ovarian cancer cells. In vivo, PACMA 31 showed tumor targeting ability and significantly suppressed ovarian tumor growth without causing toxicity to normal tissues. These irreversible small-molecule PDI inhibitors represent an important approach for the development of targeted anticancer agents for ovarian cancer therapy, and they can also serve as useful probes for investigating the biology of PDIimplicated pathways.oral bioavailability | drug resistance | BODIPY-conjugation

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“…25,26 As shown in Figure 7, the cellular uptake of M-GGLG liposomes was reduced to 55%-83% of the control group's by pretreatment with bacitracin or DTNB in HeLa, HCC1954, MDA-MB-468, and COS-7 cells, whereas the uptake of GGLG liposomes was not significantly suppressed by PDI inhibitors.…”

mentioning

confidence: 95%

Enhanced cellular uptake of maleimide-modified liposomes via thiol-mediated transport

Li¹,

Takeoka²

2014

IJN

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With a small amount of maleimide modification on the liposome surface, enhanced cellular uptake of liposomes and drug-delivery efficiency can be obtained both in vitro and in vivo. Herein, we describe the mechanisms underlying this enhanced cellular uptake. Suppression of the cellular uptake of maleimide-modified liposomes (M-GGLG, composed of 1,5-dihexadecyl N,N -diglutamyl-lysyl-L-glutamate [GGLG]/cholesterol/poly(ethylene glycol) – 1,2-distearoyl- sn -glycero-3-phosphoethanolamine [PEG 5000 -DSPE]/maleimide [M]-PEG 5000 -Glu2C 18 at a molar ratio of 5:5:0.03:0.03) caused by temperature block and addition of serum was alleviated compared with that of liposomes without maleimide modification (GGLG liposomes, composed of GGLG/cholesterol/PEG 5000 -DSPE/PEG 5000 -Glu2C 18 at a molar ratio of 5:5:0.03:0.03). When 0.01 nM N -ethylmaleimide was used to pre-block cellular thiols, the cellular uptake of M-GGLG liposomes was decreased to approximately 70% in HeLa, HCC1954, MDA-MB-468, and COS-7 cell lines. Moreover, inhibition of a thiol-related reductase such as protein disulfide isomerase resulted in a 15%–45% inhibition of the cellular uptake of M-GGLG liposomes, whereas GGLG liposomes were not influenced. Further, single and mixed inhibitors of clathrin-mediated endocytosis, caveolae-mediated endocytosis, and macropinocytosis did not efficiently inhibit the cellular uptake of M-GGLG liposomes. Using confocal microscopy, we verified that M-GGLG liposomes were localized partially in lysosomes after inhibition of the mentioned conventional endocytic pathways. Therefore, it was hypothesized that the mechanisms underlying the enhanced cellular uptake of liposomes by maleimide modification was thiol-mediated membrane trafficking, including endocytosis and energy-independent transport.

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Bacitracin inhibits the reductive activity of protein disulfide isomerase by disulfide bond formation with free cysteines in the substrate‐binding domain

Cited by 62 publications

References 29 publications

Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents

Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents

Discovery of an orally active small-molecule irreversible inhibitor of protein disulfide isomerase for ovarian cancer treatment

Enhanced cellular uptake of maleimide-modified liposomes via thiol-mediated transport

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