Structure-based discovery and in vitro validation of inhibitors of chloride intracellular channel 4 protein

Olotu, Fisayo A.; Medina‐Carmona, Encarnación; Serrano-Sanchez, Angela; Ossa, Felipe; El-Hamdaoui, Abdelaziz; Bishop, Özlem Taştan; Roldan, Jose‐Luis Ortega; Abdul-Salam, Vahitha B.

doi:10.1016/j.csbj.2022.12.040

Cited by 2 publications

(5 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CLIC3 and CLIC4 showed varying levels of residual activity in the presence of either of the three drugs, ranging from 4.1–21.5% for CLIC3, and 12.8–30% for CLIC4, respectively. These findings confirm the previous study [ 66 ] and extend the findings to indicate that all three drugs inhibit the oxidoreductase catalytic activity of CLIC1, CLIC3, and CLIC4.…”

Section: Resultssupporting

confidence: 93%

“…CLIC3 and CLIC4 showed varying levels of residual activity in the presence of either of the three drugs, ranging from 4.1-21.5% for CLIC3, and 12.8-30% for CLIC4, respectively. These findings confirm the previous study [66] and extend the findings to indicate that all three drugs inhibit the oxidoreductase catalytic activity of CLIC1, CLIC3, and CLIC4. The highly conserved and ubiquitous expression of CLIC and CLIC-like proteins across vertebrates supports the hypothesis that these proteins play critically important roles within cells [1,7,12,13,24,68,69].…”

Section: Evaluating Recombinant Clic's Enzymatic Activity Under Varyi...supporting

confidence: 92%

“…In a recent study utilizing a combination of computer-aided methods and experimental approaches, the drugs rapamycin and amphotericin B were identified as allosteric inhibitors of CLIC4, which also reversed stress-induced membrane translocation of CLIC4 and inhibited endothelial cell migration [ 66 ]. The diuretic drug IAA94 is a well-known ion channel blocker of chloride-selective ion channels, and can specifically inhibit the activity of CLIC channels when used at lower concentrations (≤100 µM) [ 67 ].…”

Section: Resultsmentioning

confidence: 99%

“…Numerous studies in the literature have shown that both protein processing and trafficking can be altered when luminal pH is changed, whether it be through posttranslational modifications and processing, misdirection, or overall changes in the integrity of organelles [54][55][56]. Furthermore, several biological processes like proteinprotein interactions [57,58], protein-ligand binding [59,60], protein interactions with In a recent study utilizing a combination of computer-aided methods and experimental approaches, the drugs rapamycin and amphotericin B were identified as allosteric inhibitors of CLIC4, which also reversed stress-induced membrane translocation of CLIC4 and inhibited endothelial cell migration [66]. The diuretic drug IAA94 is a well-known ion channel blocker of chloride-selective ion channels, and can specifically inhibit the activity of CLIC channels when used at lower concentrations (≤100 µM) [67].…”

Section: Evaluating Recombinant Clic's Enzymatic Activity Under Varyi...mentioning

confidence: 99%

See 3 more Smart Citations

In Vitro Enzymatic Studies Reveal pH and Temperature Sensitive Properties of the CLIC Proteins

Alghalayini,

Hossain,

Moghaddasi

et al. 2023

Biomolecules

View full text Add to dashboard Cite

Chloride intracellular ion channel (CLIC) proteins exist as both soluble and integral membrane proteins, with CLIC1 capable of shifting between two distinct structural conformations. New evidence has emerged indicating that members of the CLIC family act as moonlighting proteins, referring to the ability of a single protein to carry out multiple functions. In addition to their ion channel activity, CLIC family members possess oxidoreductase enzymatic activity and share significant structural and sequence homology, along with varying overlaps in their tissue distribution and cellular localization. In this study, the 2-hydroxyethyl disulfide (HEDS) assay system was used to characterize kinetic properties, as well as the temperature and pH profiles of three CLIC protein family members (CLIC1, CLIC3, CLIC4). We also assessed the effects of the drugs rapamycin and amphotericin B, on the three CLIC proteins’ enzymatic activity in the HEDS assay. Our results demonstrate CLIC1 to be highly heat-sensitive, with optimal enzymatic activity observed at neutral pH7 and at a temperature of 37 °C, while CLIC3 had higher oxidoreductase activity in more acidic pH5 and was found to be relatively heat stable. CLIC4, like CLIC1, was temperature sensitive with optimal enzymatic activity observed at 37 °C; however, it showed optimal activity in more alkaline conditions of pH8. Our current study demonstrates individual differences in the enzymatic activity between the three CLIC proteins, suggesting each CLIC protein is likely regulated in discrete ways, involving changes in the subcellular milieu and microenvironment.

show abstract

Section: Resultssupporting

confidence: 93%

Section: Evaluating Recombinant Clic's Enzymatic Activity Under Varyi...supporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Evaluating Recombinant Clic's Enzymatic Activity Under Varyi...mentioning

confidence: 99%

See 2 more Smart Citations

In Vitro Enzymatic Studies Reveal pH and Temperature Sensitive Properties of the CLIC Proteins

Alghalayini,

Hossain,

Moghaddasi

et al. 2023

Biomolecules

View full text Add to dashboard Cite

show abstract

“…Mammalian CLIC4 is an essential molecular component of cellular anion channels with poorly selective and redox-regulated activity ( Proutski et al, 2002 ; Singh and Ashley, 2007 ). Since amphotericin B and rapamycin were screened as small molecule inhibitors to CLIC4 via a high-performance computing-powered blind-docking approach ( Olotu et al, 2023 ), CLIC4 is expected to develop a novel target of clinical potential ( Leanza et al, 2013 ). However, there are fewer applications for channel-targeting insecticides.…”

Section: Discussionmentioning

confidence: 99%

Chloride intracellular channel gene knockdown induces insect cell lines death and level increases of intracellular calcium ions

Song

Yan

et al. 2023

Front. Physiol.

View full text Add to dashboard Cite

Chloride intracellular channel (CLIC) is a member of the chloride channel protein family for which growing evidence supports a pivotal role in fundamental cellular events. However, the physiological function of CLIC in insects is still rarely uncovered. The ovary-derived High Five (Hi-5) cell line isolated from the cabbage looper (Trichoplusia ni) is widely used in laboratories. Here, we studied both characteristics and functions of CLIC in Hi-5 cells (TnCLIC). We identified the TnCLIC gene in Hi-5 cells and annotated highly conserved CLIC proteins in most insect species. After RNA interference of TnCLIC, the phenomenon of significantly increased cell death suggests that the TnCLIC protein is essential for the survival of Hi-5 cells. The same lethal effect was also observed in Spodoptera frugiperda 9 and Drosophila melanogaster Schneider 2 cells after CLIC knockdown. Furthermore, we found that this kind of cell death was accompanied by increases in intracellular calcium ions after TnCLIC knockdown with the transcriptomic analyses and the detection of calcium levels. Our results provide insights into insect CLIC as a key factor for cell survival and lay the foundation for the cell death mechanism.

show abstract

Structure-based discovery and in vitro validation of inhibitors of chloride intracellular channel 4 protein

Cited by 2 publications

References 57 publications

In Vitro Enzymatic Studies Reveal pH and Temperature Sensitive Properties of the CLIC Proteins

In Vitro Enzymatic Studies Reveal pH and Temperature Sensitive Properties of the CLIC Proteins

Chloride intracellular channel gene knockdown induces insect cell lines death and level increases of intracellular calcium ions

Contact Info

Product

Resources

About