Calreticulin positively regulates the expression and function of epithelial sodium channel

Sugahara, Takuya; Koga, Tomoaki; Ueno-Shuto, Keiko; Shuto, Tsuyoshi; Watanabe, Eriko; Maekawa, Ai; Kitamura, Kenichiro; Tomita, Kimio; Mizuno, A-i; Satō, Takashi; Suico, Mary Ann; Kai, Hirofumi

doi:10.1016/j.yexcr.2009.09.023

Cited by 12 publications

(11 citation statements)

References 26 publications

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“…20) We postulate that treatment with macrolides may also inhibit overexpressionassociated constitutive ENaC activation in vitro and modulate the pathogenesis of ENaC-hyperactive lung diseases. By utilizing ENaC-overexpressing human bronchial epithelial cells (β/γENaC-16HBE14o-cells), 21) we confirmed the suppressive effect of macrolides on constitutive ENaC function. Oral treatment with azithromycin in mice harboring airway specific βENaC overexpression (C57BL/6J-βENaC-Tg mice) 7) significantly increased the forced expiratory volume in 0.1 s (FEV0.1), possibly leading to improvement of lung obstructive phenotypes such as FEV0.1%, especially at a lower dose (3 mg kg −1 ).…”

Section: Introductionsupporting

confidence: 59%

“…Cell Culture Human bronchial βγENaC-stable 16HBE14ocell line was established and grown as previously described. 21) Cells were maintained in MEM medium supplemented with 10% (v/v) fetal bovine serum (FBS), 100 IU/mL penicillin, and 100 µg/mL streptomycin. All cells were cultured in a humidified atmosphere of 5% CO 2 in air at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…We first investigated the effect of macrolide antibiotics on β/γENaC-overexpressing 16HBE14o-cells, 21) a cellular model that mimics the molecular environment of lung tissue of C57BL/6J-βENaC-Tg mice. 7) We treated β/γENaC-16HBE14ocells with serial concentration of erythromycin, clarithromycin and azithromycin, which are the major members of macrolide antibiotics.…”

Section: Macrolide Antibiotics Suppress Basal Enac Function In β/γEnamentioning

confidence: 99%

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Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation in Vitro and Modulates Downstream Pathogenesis in Vivo

Fujikawa

Kawakami

Nakashima

et al. 2020

Biological & Pharmaceutical Bulletin

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Epithelial sodium channel (ENaC) is an amiloride-sensitive sodium ion channel that is expressed in epithelial tissues. ENaC overexpression and/or hyperactivation in airway epithelial cells cause sodium overabsorption and dysregulated ciliary movement for mucus clearance; however, the agents that suppress constitutive airway ENaC activation are yet to be clinically available. Here, we focused on macrolides, which are widely used antibiotics that have many potential immunomodulatory effects. We examined whether macrolides could modulate constitutive ENaC activity and downstream events that typify cystic fibrosis (CF) and chronic obstructive pulmonary diseases (COPD) in in vitro and in vivo models of ENaC overexpression. Treatment of ENaC-overexpressing human bronchial epithelial cells (β/γENaC-16HBE14o-cells) with three macrolides (erythromycin, clarithromycin, azithromycin) confirmed dose-dependent suppression of ENaC function. For in vivo studies, mice harboring airway specific βENaC overexpression (C57BL/6J-βENaCtransgenic mice) were treated orally with azithromycin, a well-established antimicrobial agent that has been widely prescribed. Azithromycin treatment modulated pulmonary mechanics, emphysematous phenotype and pulmonary dysfunction. Notably, a lower dose (3 mg kg 1) of azithromycin significantly increased forced expiratory volume in 0.1 s (FEV0.1), an inverse indicator of bronchoconstriction. Although not statistically significant, improvement of pulmonary obstructive parameters such as emphysema and lung dysfunction (FEV0.1%) was observed. Our results demonstrate that macrolides directly attenuate constitutive ENaC function in vitro and may be promising for the treatment of obstructive lung diseases with defective mucociliary clearance, possibly by targeting ENaC hyperactivation.

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Section: Introductionsupporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

Section: Macrolide Antibiotics Suppress Basal Enac Function In β/γEnamentioning

confidence: 99%

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Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation in Vitro and Modulates Downstream Pathogenesis in Vivo

Fujikawa

Kawakami

Nakashima

et al. 2020

Biological & Pharmaceutical Bulletin

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“…Transient transfections of plasmids were performed using either Trans-IT LT1 (Mirus, Madison, WI) or Hilymax (Dojin Co. Ltd, Tokyo, Japan) by following the manufacturer’s recommendations as described earlier [13,14]. For the transfection of si-RNAs, Trans-IT TKO (Mirus) was used as previously described [15]. Fifty nanomolar of si-Derlin-1 and Stealth ™ RNAi negative Universal Control (control siRNA) were transfected into 90% confluent cells to knockdown Derlin-1.…”

Section: Methodsmentioning

confidence: 99%

Posttranslational negative regulation of glycosylated and non-glycosylated BCRP expression by Derlin-1

Sugiyama

Shuto

Suzuki

et al. 2011

Biochemical and Biophysical Research Communications

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Human breast cancer resistance protein (BCRP)/MXR/ABCG2 is a well-recognized ABC half-transporter that is highly expressed at the apical membrane of many normal tissues and cancer cells. BCRP facilitates disposition of endogenous and exogenous harmful xenobiotics to protect cells/tissues from xenobiotic-induced toxicity. Despite the enormous impact of BCRP in the physiological and pathophysiological regulation of the transport of a wide variety of substrates, little is known about the factors that regulate posttranslational expression of BCRP. Here, we identified Derlin-1, a member of a family of proteins that bears homology to yeast Der1p, as a posttranslational regulator of BCRP expression. Overexpression of Derlin-1 suppressed ER to Golgi transport of wild-type (WT) BCRP that is known to be efficiently trafficked to the plasma membrane. On the other hand, protein expression of N596Q variant of BCRP, N-linked glycosylation- deficient mutant that preferentially undergoes ubiquitin-mediated ER-associated degradation (ERAD), was strongly suppressed by the overexpression of Derlin-1, whereas knockdown of Derlin-1 stabilized N596Q protein, suggesting a negative regulatory role of Derlin-1 for N596Q protein expression. Notably, knockdown of Derlin-1 also stabilized the expression of tunicamycin-induced deglycosylated WT BCRP protein, implying the importance of glycosylation state for the recognition of BCRP by Derlin- 1. Thus, our data demonstrate that Derlin-1 is a negative regulator for both glycosylated and non-glycosylated BCRP expression and provide a novel posttranslational regulatory mechanism of BCRP by Derlin-1.

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“…Mouse embryonic fibroblasts deficient in calreticulin exhibited increased ER retention of collagen, the extracellular matrix component [ 22 ]. Calreticulin up-regulates the expression of epithelial sodium channel (ENaC) subunits, and increased channel activities were observed in cells overexpressing calreticulin [ 23 ]. Calreticulin has also been shown to stabilize and promote the stability of human insulin receptors in the ER [ 24 ].…”

Section: Calreticulin In the Ermentioning

confidence: 99%

Calreticulin: Roles in Cell-Surface Protein Expression

2014

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In order to perform their designated functions, proteins require precise subcellular localizations. For cell-surface proteins, such as receptors and channels, they are able to transduce signals only when properly targeted to the cell membrane. Calreticulin is a multi-functional chaperone protein involved in protein folding, maturation, and trafficking. However, evidence has been accumulating that calreticulin can also negatively regulate the surface expression of certain receptors and channels. In these instances, depletion of calreticulin enhances cell-surface expression and function. In this review, we discuss the role of calreticulin with a focus on its negative effects on the expression of cell-surface proteins.

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Calreticulin positively regulates the expression and function of epithelial sodium channel

Cited by 12 publications

References 26 publications

Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation <i>in Vitro</i> and Modulates Downstream Pathogenesis <i>in Vivo</i>

Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation <i>in Vitro</i> and Modulates Downstream Pathogenesis <i>in Vivo</i>

Posttranslational negative regulation of glycosylated and non-glycosylated BCRP expression by Derlin-1

Calreticulin: Roles in Cell-Surface Protein Expression

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