Document
type: Class

Continuant
type: Class

Entity
type: Class

PKC-Mediated Orai1 Channel Phosphorylation Modulates Ca2%2b Signaling in HeLa Cells
date time value: article date 9927
accesstype: All Open Access, Gold, Green
most specific type: Document
freetext keyword: phosphomimetic; phosphorylated Orai1; PKC; SOCE adenosine triphosphate; calcium ion; calcium release activated calcium channel 1; inositol 1 4 5 trisphosphate receptor type 2; inositol 1 4 5 trisphosphate receptor; protein kinase C; receptor protein; serine; stromal interaction molecule 1; thapsigargin; unclassified drug; adenosine triphosphate; calcium; calcium channel; calcium release activated calcium channel 1; ORAI1 protein, human; thapsigargin; Article; bicinchoninic acid assay; bioassay; calcium depletion; calcium signaling; calcium transport; confocal microscopy; controlled study; endoplasmic reticulum calcium depletion; gene overexpression; genetic transfection; HeLa cell line; human; human cell; immunoprecipitation; point mutation; polymerase chain reaction; protein kinase c mediated orai1 channel phosphorylation; protein phosphorylation; proximity ligation assay; Sanger sequencing; spectrofluorometry; store operated calcium entry; Western blotting; HeLa cell line; metabolism; phosphorylation; Adenosine Triphosphate; Calcium; Calcium Channels; HeLa Cells; Humans; ORAI1 Protein; Phosphorylation; Thapsigargin
issue: 13
type: Article
type: Entity
estado: published
doi: 10.3390/cells11132037
abstract: The overexpression of the Orai1 channel inhibits SOCE when using the Ca2%2b readdition protocol. However, we found that HeLa cells overexpressing the Orai1 channel displayed enhanced Ca2%2b entry and a limited ER depletion in response to the combination of ATP and thapsigargin (TG) in the presence of external Ca2%2b . As these effects require the combination of an agonist and TG, we decided to study whether the phosphorylation of Orai1 S27/S30 residues had any role using two different mutants: Orai1-S27/30A (O1-AA, phosphorylation-resistant) and Orai1-S27/30D (O1-DD, phosphomimetic). Both O1-wt and O1-AA supported enhanced Ca2%2b entry, but this was not the case with O1-E106A (dead-pore mutant), O1-DD, and O1-AA-E106A, while O1-wt, O1-E106A, and O1-DD inhibited the ATP and TG-induced reduction of ER [Ca2%2b], suggesting that the phosphorylation of O1 S27/30 interferes with the IP3R activity. O1-wt and O1-DD displayed an increased interaction with IP3R in response to ATP and TG; however, the O1-AA channel decreased this interaction. The expression of mCherry-O1-AA increased the frequency of ATP-induced sinusoidal [Ca2%2b]i oscillations, while mCherry-O1-wt and mCherry-O1-DD decreased this frequency. These data suggest that the combination of ATP and TG stimulates Ca2%2b entry, and the phosphorylation of Orai1 S27/30 residues by PKC reduces IP3R-mediated Ca2%2b release. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
volume: 11
tradenames: C-DiGit, LI COR, United States; GeneJET, Thermo
most specific type: Article
has funding vehicle: funding3859
type: Document
e iscopus: 2-s2.0-85132807613
type: Generically Dependent Continuant
type: Continuant
manufactures: Carl Zeiss; Nikon, Netherlands; LI COR, United States; Thermo
pmid: 35805121
has publication venue: Cells
origen: scopus
artnum: 2037
type: Information Content Entity
related by: authorship_68912
type: Thing
chemicalscas: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; calcium ion, 14127-61-8; protein kinase C, 141436-78-4; protein kinase C theta; serine, 56-45-1, 6898-95-9; serine threonine protein kinase D2; thapsigargin, 67526-95-8; calcium, 7440-70-2, 14092-94-5; Adenosine Triphosphate; Calcium; Calcium Channels; ORAI1 Protein; ORAI1 protein, human; Thapsigargin
crisid: 9927

Generically Dependent Continuant
type: Class

RDF description of PKC-Mediated Orai1 Channel Phosphorylation Modulates Ca2%2b Signaling in HeLa Cells - http://orbis.uaslp.mx/vivo/individual/scopus9926
rights: termsOfUse
date: 2026-08-28T02:11:28
publisher: vivo
type: Document

Article
type: Class

Information Content Entity
type: Class

funding3859
type: Relationship
type: BFO_0000020
type: Entity
type: Thing
type: Grant
type: Continuant
label: Texas A%26M-Conacyt Collaborative Research Grant Program 19-20-052; Consejo Nacional de Ciencia y Tecnología, CONACYT: 338030, PID2019-104084 GB-C21; European Regional Development Fund, ERDF: FC 2016-2803, GR18061; Agencia Estatal de Investigación, AEI
funding vehicle for: PKC-Mediated Orai1 Channel Phosphorylation Modulates Ca2%2b Signaling in HeLa Cells

Cells
type: Information Content Entity
type: Continuant
type: Entity
type: Collection
type: Periodical
type: Document
type: Journal
publication venue for: PKC-Mediated Orai1 Channel Phosphorylation Modulates Ca2%2b Signaling in HeLa Cells
type: Thing
type: Generically Dependent Continuant