Inhibition of nitric oxide‐activated guanylyl cyclase by calmodulin antagonists

James, LR; Griffiths, C. H.; Garthwaite, John; Bellamy, Tomas C.

doi:10.1111/j.1476-5381.2009.00416.x

Cited by 11 publications

(10 citation statements)

References 58 publications

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“…CyaA and sarcoplasmatic/endoplasmatic reticulum Ca 2+ -ATPase (SERCA)), where CDZ acts as a potent inhibitor, it was suggested that the potency of inhibition is correlated with the hydrophobicity and the size of the inhibitor [ 21 , 49 ]. In contrast to most of above CaM-inhibitors, CDZ inhibits many CaM-targets in a CaM-independent manner [ 18 , 21 , 49 – 52 ]. Table 1 summarizes the effects of CaM-inhibitors on CaM-target interactions.…”

Section: Introductionmentioning

confidence: 99%

Effects of 39 Compounds on Calmodulin-Regulated Adenylyl Cyclases AC1 and Bacillus anthracis Edema Factor

Lübker

Seifert

2015

PLoS ONE

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Adenylyl cyclases (ACs) catalyze the conversion of ATP into the second messenger cAMP. Membranous AC1 (AC1) is involved in processes of memory and learning and in muscle pain. The AC toxin edema factor (EF) of Bacillus anthracis is involved in the development of anthrax. Both ACs are stimulated by the eukaryotic Ca2+-sensor calmodulin (CaM). The CaM-AC interaction could constitute a potential target to enhance or impair the AC activity of AC1 and EF to intervene in above (patho)physiological mechanisms. Thus, we analyzed the impact of 39 compounds including typical CaM-inhibitors, an anticonvulsant, an anticholinergic, antidepressants, antipsychotics and Ca2+-antagonists on CaM-stimulated catalytic activity of AC1 and EF. Compounds were tested at 10 μM, i.e., a concentration that can be reached therapeutically for certain antidepressants and antipsychotics. Calmidazolium chloride decreased CaM-stimulated AC1 activity moderately by about 30%. In contrast, CaM-stimulated EF activity was abrogated by calmidazolium chloride and additionally decreased by chlorpromazine, felodipine, penfluridol and trifluoperazine by about 20–40%. The activity of both ACs was decreased by calmidazolium chloride in the presence and absence of CaM. Thus, CaM-stimulated AC1 activity is more insensitive to inhibition by small molecules than CaM-stimulated EF activity. Inhibition of AC1 and EF by calmidazolium chloride is largely mediated via a CaM-independent allosteric mechanism.

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Section: Introductionmentioning

confidence: 99%

Effects of 39 Compounds on Calmodulin-Regulated Adenylyl Cyclases AC1 and Bacillus anthracis Edema Factor

Lübker

Seifert

2015

PLoS ONE

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“…However, we detected no manipulation of AMPylation by any of the tested compounds relative to the DMSO control ( Figure S5 ). The activity of calmidazolium against HYPE and VopS is likely attributable to its vast biological promiscuity, rather than some Fic-specific mechanism [ 38 , 39 , 40 ].…”

Section: Resultsmentioning

confidence: 99%

A Fluorescence Polarization-Based High-Throughput Screen to Identify the First Small-Molecule Modulators of the Human Adenylyltransferase HYPE/FICD

Camara

George

Hebner

et al. 2020

IJMS

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The covalent transfer of the AMP portion of ATP onto a target protein—termed adenylylation or AMPylation—by the human Fic protein HYPE/FICD has recently garnered attention as a key regulatory mechanism in endoplasmic reticulum homeostasis, neurodegeneration, and neurogenesis. As a central player in such critical cellular events, high-throughput screening (HTS) efforts targeting HYPE-mediated AMPylation warrant investigation. Herein, we present a dual HTS assay for the simultaneous identification of small-molecule activators and inhibitors of HYPE AMPylation. Employing the fluorescence polarization of an ATP analog fluorophore—Fl-ATP—we developed and optimized an efficient, robust assay that monitors HYPE autoAMPylation and is amenable to automated, high-throughput processing of diverse chemical libraries. Challenging our pilot screen with compounds from the LOPAC, Spectrum, MEGx, and NATx libraries yielded 0.3% and 1% hit rates for HYPE activators and inhibitors, respectively. Further, these hits were assessed for dose-dependency and validated via orthogonal biochemical AMPylation assays. We thus present a high-quality HTS assay suitable for tracking HYPE’s enzymatic activity, and the resultant first small-molecule manipulators of HYPE-promoted autoAMPylation.

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“…Both CALMID and TFP were previously shown to inhibit CaM activity primarily by binding directly to the protein (Matsushima et al, 2000;Sunagawa et al, 2000). However, CALMID and TFP probably exert many of their actions not only via their binding to CaM, but also by interfering directly with a number of upstream (Qin et al, 2009) or downstream targets of CaM signaling (James et al, 2009;Sunagawa et al, 2000). For example, the Rho family GTPases, e.g.…”

Section: Discussionmentioning

confidence: 99%

Calmodulin inhibition regulates morphological and functional changes related to the actin cytoskeleton in pure microglial cells

Szabó

Dulka

Gulya

2016

Brain Research Bulletin

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The roles of calmodulin (CaM), a multifunctional intracellular calcium receptor protein, as concerns selected morphological and functional characteristics of pure microglial cells derived from mixed primary cultures from embryonal forebrains of rats, were investigated through use of the CaM antagonists calmidazolium (CALMID) and trifluoperazine (TFP). The intracellular localization of the CaM protein relative to phalloidin, a bicyclic heptapeptide that binds only to filamentous actin, and the ionized calcium-binding adaptor molecule 1 (Iba1), a microglia-specific actin-binding protein, was determined by immunocytochemistry, with quantitative analysis by immunoblotting. In unchallenged and untreated (control) microglia, high concentrations of CaM protein were found mainly perinuclearly in ameboid microglia, while the cell cortex had a smaller CaM content that diminished progressively deeper into the branches in the ramified microglia. The amounts and intracellular distributions of both Iba1 and CaM proteins were altered after lipopolysaccharide (LPS) challenge in activated microglia. CALMID and TFP exerted different, sometimes opposing, effects on many morphological, cytoskeletal and functional characteristics of the microglial cells. They affected the CaM and Iba1 protein expressions and their intracellular localizations differently, inhibited cell proliferation, viability and fluid-phase phagocytosis to different degrees both in unchallenged and in LPS-treated (immunologically challenged) cells, and differentially affected the reorganization of the actin cytoskeleton in the microglial cell cortex, influencing lamellipodia, filopodia and podosome formation. In summary, these CaM antagonists altered different aspects of filamentous actin-based cell morphology and related functions with variable efficacy, which could be important in deciphering the roles of CaM in regulating microglial functions in health and disease.

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Inhibition of nitric oxide‐activated guanylyl cyclase by calmodulin antagonists

Cited by 11 publications

References 58 publications

Effects of 39 Compounds on Calmodulin-Regulated Adenylyl Cyclases AC1 and Bacillus anthracis Edema Factor

Effects of 39 Compounds on Calmodulin-Regulated Adenylyl Cyclases AC1 and Bacillus anthracis Edema Factor

A Fluorescence Polarization-Based High-Throughput Screen to Identify the First Small-Molecule Modulators of the Human Adenylyltransferase HYPE/FICD

Calmodulin inhibition regulates morphological and functional changes related to the actin cytoskeleton in pure microglial cells

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