Bafilomycin and Concanamycin Derivatives as Specific Inhibitors of P- and V-Type ATPases

Dröse, Stefan; Bindseil, Kai U.; Bowman, Emma Jean; Zeeck, Axel; Altendorf, Karlheinz

doi:10.1007/978-3-642-72511-1_66

Cited by 189 publications

(225 citation statements)

References 6 publications

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“…bafilomycin A1 (Baf) or concanamycin A (ConcA), which have been shown to be potent and selective inhibitors of the vacuolar H þ -ATPase [Drose and Altendorf, 1997]. As expected, pH G was severely alkalanized to values that were similar to those of the cytoplasm [Llopis et al, 1998] (Table II).…”

Section: Depolymerization But Not Stabilization Of F-actin Raises Intmentioning

confidence: 60%

Actin filaments are involved in the maintenance of Golgi cisternae morphology and intra‐Golgi pH

Lázaro‐Diéguez

Jiménez

Barth

et al. 2006

Cell Motil. Cytoskeleton

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Here we examine the contribution of actin dynamics to the architecture and pH of the Golgi complex. To this end, we have used toxins that depolymerize (cytochalasin D, latrunculin B, mycalolide B, and Clostridium botulinum C2 toxin) or stabilize (jasplakinolide) filamentous actin. When various clonal cell lines were examined by epifluorescence microscopy, all of these actin toxins induced compaction of the Golgi complex. However, ultrastructural analysis by transmission electron microscopy and electron tomography/three-dimensional modelling of the Golgi complex showed that F-actin depolymerization first induces perforation/fragmentation and severe swelling of Golgi cisternae, which leads to a completely disorganized structure. In contrast, F-actin stabilization results only in cisternae perforation/fragmentation. Concomitantly to actin depolymerization-induced cisternae swelling and disorganization, the intra-Golgi pH significantly increased. Similar ultrastructural and Golgi pH alkalinization were observed in cells treated with the vacuolar H þ -ATPases inhibitors bafilomycin A1 and concanamycin A. Overall, these results suggest that actin filaments are implicated in the preservation of the flattened shape of Golgi cisternae. This maintenance seems to be mediated by the regulation of the state of F-actin assembly on the Golgi pH homeostasis. Cell Motil. Cytoskeleton 63: 778-791, 2006. '

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Section: Depolymerization But Not Stabilization Of F-actin Raises Intmentioning

confidence: 60%

Actin filaments are involved in the maintenance of Golgi cisternae morphology and intra‐Golgi pH

Lázaro‐Diéguez

Jiménez

Barth

et al. 2006

Cell Motil. Cytoskeleton

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“…4). Ammonium chloride (NH 4 Cl) at 5 mM, which dissipates the proton gradient over the membrane, reduced the ABA-GE uptake activity by 28%, and 0.5 mM bafilomycin A1, a vacuolar proton pump (V-ATPase) inhibitor (Dröse and Altendorf, 1997), reduced it by 43%. Residual proton gradients present in isolated vacuoles may energize transport even when V-ATPases are inhibited.…”

Section: Enzymatic Synthesis Of Radiolabeled Aba-gementioning

confidence: 99%

Vacuolar Transport of Abscisic Acid Glucosyl Ester Is Mediated by ATP-Binding Cassette and Proton-Antiport Mechanisms in Arabidopsis

et al. 2013

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Abscisic acid (ABA) is a key plant hormone involved in diverse physiological and developmental processes, including abiotic stress responses and the regulation of stomatal aperture and seed germination. Abscisic acid glucosyl ester (ABA-GE) is a hydrolyzable ABA conjugate that accumulates in the vacuole and presumably also in the endoplasmic reticulum. Deconjugation of ABA-GE by the endoplasmic reticulum and vacuolar b-glucosidases allows the rapid formation of free ABA in response to abiotic stress conditions such as dehydration and salt stress. ABA-GE further contributes to the maintenance of ABA homeostasis, as it is the major ABA catabolite exported from the cytosol. In this work, we identified that the import of ABA-GE into vacuoles isolated from Arabidopsis (Arabidopsis thaliana) mesophyll cells is mediated by two distinct membrane transport mechanisms: proton gradientdriven and ATP-binding cassette (ABC) transporters. Both systems have similar K m values of approximately 1 mM. According to our estimations, this low affinity appears nevertheless to be sufficient for the continuous vacuolar sequestration of ABA-GE produced in the cytosol. We further demonstrate that two tested multispecific vacuolar ABCC-type ABC transporters from Arabidopsis exhibit ABA-GE transport activity when expressed in yeast (Saccharomyces cerevisiae), which also supports the involvement of ABC transporters in ABA-GE uptake. Our findings suggest that the vacuolar ABA-GE uptake is not mediated by specific, but rather by several, possibly multispecific, transporters that are involved in the general vacuolar sequestration of conjugated metabolites.

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“…Third, we blocked vesicle reacidification by bafilomycin A1, a specific inhibitor of the V-ATPase on secretory vesicles ( Dröse and Altendorf, 1997). This should abolish transient events caused by reacidification, but not those that reflect complete release and cargo diffusion.…”

Section: Transient Events Represent Complete Release As Well As Retrimentioning

confidence: 99%

Matrix-Dependent Local Retention of Secretory Vesicle Cargo in Cortical Neurons

Wit¹,

Toonen²,

Verhage³

2009

J. Neurosci.

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Neurons secrete many diffusible signals from synaptic and other secretory vesicles. We characterized secretion of guidance cues, neuropeptides, neurotrophins, and proteases from single secretory vesicles using pHluorin-tagged cargo in cortical neurons. Stimulation triggered transient and persistent fusion events. Transient events represented full release followed by cargo diffusion or incomplete release followed by vesicle retrieval, as previously observed in neuroendocrine cells. Unexpectedly, we also observed that certain cargo, such as Semaphorin 3A (Sema3A), was delivered at the cell surface as stable deposits. Stable deposits and transient events were observed for single cargo and both were SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) and calcium dependent. The ratio between stable and transient events did not depend on cargo size, subcellular localization (synaptic vs extrasynaptic secretion), or the presence of the extracellular matrix. Instead, the ratio is cargo specific and depends on an interaction with the vesicle matrix through a basic domain in the cargo protein. Inhibition of this interaction through deletion of the basic domain in Sema3A abolished stable deposits and rendered all events transient. Strikingly, cargo favoring transient release was stably deposited after corelease with cargo favoring stable deposit. These data argue against cargo diffusion after exocytosis as a general principle. Instead, the vesicle matrix retains secreted signals, probably for focal signaling at the cell surface.

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Bafilomycin and Concanamycin Derivatives as Specific Inhibitors of P- and V-Type ATPases

Cited by 189 publications

References 6 publications

Actin filaments are involved in the maintenance of Golgi cisternae morphology and intra‐Golgi pH

Actin filaments are involved in the maintenance of Golgi cisternae morphology and intra‐Golgi pH

Vacuolar Transport of Abscisic Acid Glucosyl Ester Is Mediated by ATP-Binding Cassette and Proton-Antiport Mechanisms in Arabidopsis

Matrix-Dependent Local Retention of Secretory Vesicle Cargo in Cortical Neurons

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