Korean J Physiol Pharmacol.  2018 May;22(3):343-348. 10.4196/kjpp.2018.22.3.343.

Increased store-operated Ca²⁺ entry mediated by GNB5 and STIM1

• Kang N
• Kang JY
• Park S
• Shin DM

• KMID: 2410098
• DOI: http://doi.org/10.4196/kjpp.2018.22.3.343

Abstract

Recent human genetic studies have shown that Gβ5 is related to various clinical symptoms, such as sinus bradycardia, cognitive disability, and attention deficit hyperactivity disorder. Although the calcium signaling cascade is closely associated with a heterotrimeric G-protein, the function of Gβ5 in calcium signaling and its relevance to clinical symptoms remain unknown. In this study, we investigated the in vitro changes of store-operated calcium entry (SOCE) with exogenous expression of Gβ5. The cells expressing Gβ5 had enhanced SOCE after depletion of calcium ion inside the endoplasmic reticulum. Gβ5 also augmented Stim1- and Orai1-dependent SOCE. An ORAI1 loss-of-function mutant did not show inhibition of Gβ5-induced SOCE, and a STIM1-ERM truncation mutant showed no enhancement of SOCE. These results suggested a novel role of GNB5 and Stim1, and provided insight into the regulatory mechanism of SOCE.

Keyword

Ca²⁺ signaling; GNB5; Orai1; STIM1; Store-operated Ca²⁺ entry

MeSH Terms

Attention Deficit Disorder with Hyperactivity
Bradycardia
Calcium
Calcium Signaling
Endoplasmic Reticulum
GTP-Binding Proteins
Humans
In Vitro Techniques
Calcium
GTP-Binding Proteins

Figure

• 

  Fig. 1 Enhanced store-operated Ca2+ entry induced by overexpression of GNB5After depletion of ER Ca2+ stores using cyclopiazonic acid and EGTA, Ca2+ entry was measure by Fura2-AM fluorescent signal ratios. A representative trace (A) summarizes the data (B), and the number inside the column is the total cell number from multiple independent experiments (n=3-4). Data were expressed as the mean±SEM. *p<0.05 compared with GFP.

• 

  Fig. 2 Increased Stim1-Orai1 dependent store-operated Ca2+ entry after GNB5 overexpression.Gβ5, Stim1, and Orai1 were co-expressed in HEK293T cells. A representative trace (A) summarizes the data (B), and the number inside the column is the total cell number from multiple independent experiments (n=3-4). Data were expressed as the mean±SEM. *p<0.05 compared with control.

• 

  Fig. 3 Effects of a loss-of-functional Orai1 mutant on store-operated Ca2+ entry.Wild-type Gβ5, wild-type Stim1, and Orai1R91W were co-expressed in HEK293T cells. A representative trace (A) summarizes the data (B), and the number inside the column is the total cell number from multiple independent experiments (n=3-4). Data were expressed as the mean±SEM. *p<0.05 compared with control.

• 

  Fig. 4 Diminished enhancement of the store-operated Ca2+ entry induced by Gβ5 and the co-expression of a dominant-negative Stim1.Wild-type Gβ5, ΔERM-Stim1D76A, and wild-type Orai1 were co-expressed in HEK293T cells. A representative trace (A) summarizes the data (B), and the number inside the column is the total cell number from multiple independent experiments (n=3-4). N. D.=not determined.

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