Trpm2 enhances physiological bioenergetics and protects against pathological oxidative cardiac injury: Role of Pyk2 phosphorylation

Barbara Miller, Ju Fang Wang, Jianliang Song, Xue Qian Zhang, Iwona Hirschler-Laszkiewicz, Shanmughapriya Santhanam, Dhanendra Tomar, Sudasan Rajan, Arthur M. Feldman, Muniswamy Madesh, Shey Shing Sheu, Joseph Y. Cheung

Research output: Contribution to journalArticle

Abstract

The mechanisms by which Trpm2 channels enhance mitochondrial bioenergetics and protect against oxidative stress-induced cardiac injury remain unclear. Here, the role of proline-rich tyrosine kinase 2 (Pyk2) in Trpm2 signaling is explored. Activation of Trpm2 in adult myocytes with H2O2 resulted in 10- to 21-fold increases in Pyk2 phosphorylation in wild-type (WT) myocytes which was significantly lower (~40%) in Trpm2 knockout (KO) myocytes. Pyk2 phosphorylation was inhibited (~54%) by the Trpm2 blocker clotrimazole. Buffering Trpm2-mediated Ca2+ increase with 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) resulted in significantly reduced pPyk2 in WT but not in KO myocytes, indicating Ca2+ influx through activated Trpm2 channels phosphorylated Pyk2. Part of phosphorylated Pyk2 translocated from cytosol to mitochondria which has been previously shown to augment mitochondrial Ca2+ uptake and enhance adenosine triphosphate generation. Although Trpm2-mediated Ca2+ influx phosphorylated Ca2+-calmodulin kinase II (CaMKII), the CaMKII inhibitor KN93 did not significantly affect Pyk2 phosphorylation in H2O2-treated WT myocytes. After ischemia/reperfusion (I/R), Pyk2 phosphorylation and its downstream prosurvival signaling molecules (pERK1/2 and pAkt) were significantly lower in KO-I/R when compared with WT-I/R hearts. After hypoxia/reoxygenation, mitochondrial membrane potential was lower and superoxide level was higher in KO myocytes, and were restored to WT values by the mitochondria-targeted superoxide scavenger MitoTempo. Our results suggested that Ca2+ influx via tonically activated Trpm2 phosphorylated Pyk2, part of which translocated to mitochondria, resulting in better mitochondrial bioenergetics to maintain cardiac health. After I/R, Pyk2 activated prosurvival signaling molecules and prevented excessive increases in reactive oxygen species, thereby affording protection from I/R injury.

Original languageEnglish (US)
Pages (from-to)15048-15060
Number of pages13
JournalJournal of Cellular Physiology
Volume234
Issue number9
DOIs
StatePublished - Sep 1 2019

Fingerprint

Focal Adhesion Kinase 2
Phosphorylation
Energy Metabolism
Muscle Cells
Wounds and Injuries
Mitochondria
Reperfusion
Ischemia
Calcium-Calmodulin-Dependent Protein Kinases
Superoxides
Clotrimazole
Molecules
Ethane
Oxidative stress
Mitochondrial Membrane Potential
Reperfusion Injury
Cytosol
Reactive Oxygen Species
Oxidative Stress
Adenosine Triphosphate

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Miller, Barbara ; Wang, Ju Fang ; Song, Jianliang ; Zhang, Xue Qian ; Hirschler-Laszkiewicz, Iwona ; Santhanam, Shanmughapriya ; Tomar, Dhanendra ; Rajan, Sudasan ; Feldman, Arthur M. ; Madesh, Muniswamy ; Sheu, Shey Shing ; Cheung, Joseph Y. / Trpm2 enhances physiological bioenergetics and protects against pathological oxidative cardiac injury : Role of Pyk2 phosphorylation. In: Journal of Cellular Physiology. 2019 ; Vol. 234, No. 9. pp. 15048-15060.
@article{275141313979427b9cc7e68b160c5516,
title = "Trpm2 enhances physiological bioenergetics and protects against pathological oxidative cardiac injury: Role of Pyk2 phosphorylation",
abstract = "The mechanisms by which Trpm2 channels enhance mitochondrial bioenergetics and protect against oxidative stress-induced cardiac injury remain unclear. Here, the role of proline-rich tyrosine kinase 2 (Pyk2) in Trpm2 signaling is explored. Activation of Trpm2 in adult myocytes with H2O2 resulted in 10- to 21-fold increases in Pyk2 phosphorylation in wild-type (WT) myocytes which was significantly lower (~40{\%}) in Trpm2 knockout (KO) myocytes. Pyk2 phosphorylation was inhibited (~54{\%}) by the Trpm2 blocker clotrimazole. Buffering Trpm2-mediated Ca2+ increase with 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) resulted in significantly reduced pPyk2 in WT but not in KO myocytes, indicating Ca2+ influx through activated Trpm2 channels phosphorylated Pyk2. Part of phosphorylated Pyk2 translocated from cytosol to mitochondria which has been previously shown to augment mitochondrial Ca2+ uptake and enhance adenosine triphosphate generation. Although Trpm2-mediated Ca2+ influx phosphorylated Ca2+-calmodulin kinase II (CaMKII), the CaMKII inhibitor KN93 did not significantly affect Pyk2 phosphorylation in H2O2-treated WT myocytes. After ischemia/reperfusion (I/R), Pyk2 phosphorylation and its downstream prosurvival signaling molecules (pERK1/2 and pAkt) were significantly lower in KO-I/R when compared with WT-I/R hearts. After hypoxia/reoxygenation, mitochondrial membrane potential was lower and superoxide level was higher in KO myocytes, and were restored to WT values by the mitochondria-targeted superoxide scavenger MitoTempo. Our results suggested that Ca2+ influx via tonically activated Trpm2 phosphorylated Pyk2, part of which translocated to mitochondria, resulting in better mitochondrial bioenergetics to maintain cardiac health. After I/R, Pyk2 activated prosurvival signaling molecules and prevented excessive increases in reactive oxygen species, thereby affording protection from I/R injury.",
author = "Barbara Miller and Wang, {Ju Fang} and Jianliang Song and Zhang, {Xue Qian} and Iwona Hirschler-Laszkiewicz and Shanmughapriya Santhanam and Dhanendra Tomar and Sudasan Rajan and Feldman, {Arthur M.} and Muniswamy Madesh and Sheu, {Shey Shing} and Cheung, {Joseph Y.}",
year = "2019",
month = "9",
day = "1",
doi = "10.1002/jcp.28146",
language = "English (US)",
volume = "234",
pages = "15048--15060",
journal = "Journal of Cellular Physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "9",

}

Trpm2 enhances physiological bioenergetics and protects against pathological oxidative cardiac injury : Role of Pyk2 phosphorylation. / Miller, Barbara; Wang, Ju Fang; Song, Jianliang; Zhang, Xue Qian; Hirschler-Laszkiewicz, Iwona; Santhanam, Shanmughapriya; Tomar, Dhanendra; Rajan, Sudasan; Feldman, Arthur M.; Madesh, Muniswamy; Sheu, Shey Shing; Cheung, Joseph Y.

In: Journal of Cellular Physiology, Vol. 234, No. 9, 01.09.2019, p. 15048-15060.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Trpm2 enhances physiological bioenergetics and protects against pathological oxidative cardiac injury

T2 - Role of Pyk2 phosphorylation

AU - Miller, Barbara

AU - Wang, Ju Fang

AU - Song, Jianliang

AU - Zhang, Xue Qian

AU - Hirschler-Laszkiewicz, Iwona

AU - Santhanam, Shanmughapriya

AU - Tomar, Dhanendra

AU - Rajan, Sudasan

AU - Feldman, Arthur M.

AU - Madesh, Muniswamy

AU - Sheu, Shey Shing

AU - Cheung, Joseph Y.

PY - 2019/9/1

Y1 - 2019/9/1

N2 - The mechanisms by which Trpm2 channels enhance mitochondrial bioenergetics and protect against oxidative stress-induced cardiac injury remain unclear. Here, the role of proline-rich tyrosine kinase 2 (Pyk2) in Trpm2 signaling is explored. Activation of Trpm2 in adult myocytes with H2O2 resulted in 10- to 21-fold increases in Pyk2 phosphorylation in wild-type (WT) myocytes which was significantly lower (~40%) in Trpm2 knockout (KO) myocytes. Pyk2 phosphorylation was inhibited (~54%) by the Trpm2 blocker clotrimazole. Buffering Trpm2-mediated Ca2+ increase with 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) resulted in significantly reduced pPyk2 in WT but not in KO myocytes, indicating Ca2+ influx through activated Trpm2 channels phosphorylated Pyk2. Part of phosphorylated Pyk2 translocated from cytosol to mitochondria which has been previously shown to augment mitochondrial Ca2+ uptake and enhance adenosine triphosphate generation. Although Trpm2-mediated Ca2+ influx phosphorylated Ca2+-calmodulin kinase II (CaMKII), the CaMKII inhibitor KN93 did not significantly affect Pyk2 phosphorylation in H2O2-treated WT myocytes. After ischemia/reperfusion (I/R), Pyk2 phosphorylation and its downstream prosurvival signaling molecules (pERK1/2 and pAkt) were significantly lower in KO-I/R when compared with WT-I/R hearts. After hypoxia/reoxygenation, mitochondrial membrane potential was lower and superoxide level was higher in KO myocytes, and were restored to WT values by the mitochondria-targeted superoxide scavenger MitoTempo. Our results suggested that Ca2+ influx via tonically activated Trpm2 phosphorylated Pyk2, part of which translocated to mitochondria, resulting in better mitochondrial bioenergetics to maintain cardiac health. After I/R, Pyk2 activated prosurvival signaling molecules and prevented excessive increases in reactive oxygen species, thereby affording protection from I/R injury.

AB - The mechanisms by which Trpm2 channels enhance mitochondrial bioenergetics and protect against oxidative stress-induced cardiac injury remain unclear. Here, the role of proline-rich tyrosine kinase 2 (Pyk2) in Trpm2 signaling is explored. Activation of Trpm2 in adult myocytes with H2O2 resulted in 10- to 21-fold increases in Pyk2 phosphorylation in wild-type (WT) myocytes which was significantly lower (~40%) in Trpm2 knockout (KO) myocytes. Pyk2 phosphorylation was inhibited (~54%) by the Trpm2 blocker clotrimazole. Buffering Trpm2-mediated Ca2+ increase with 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) resulted in significantly reduced pPyk2 in WT but not in KO myocytes, indicating Ca2+ influx through activated Trpm2 channels phosphorylated Pyk2. Part of phosphorylated Pyk2 translocated from cytosol to mitochondria which has been previously shown to augment mitochondrial Ca2+ uptake and enhance adenosine triphosphate generation. Although Trpm2-mediated Ca2+ influx phosphorylated Ca2+-calmodulin kinase II (CaMKII), the CaMKII inhibitor KN93 did not significantly affect Pyk2 phosphorylation in H2O2-treated WT myocytes. After ischemia/reperfusion (I/R), Pyk2 phosphorylation and its downstream prosurvival signaling molecules (pERK1/2 and pAkt) were significantly lower in KO-I/R when compared with WT-I/R hearts. After hypoxia/reoxygenation, mitochondrial membrane potential was lower and superoxide level was higher in KO myocytes, and were restored to WT values by the mitochondria-targeted superoxide scavenger MitoTempo. Our results suggested that Ca2+ influx via tonically activated Trpm2 phosphorylated Pyk2, part of which translocated to mitochondria, resulting in better mitochondrial bioenergetics to maintain cardiac health. After I/R, Pyk2 activated prosurvival signaling molecules and prevented excessive increases in reactive oxygen species, thereby affording protection from I/R injury.

UR - http://www.scopus.com/inward/record.url?scp=85059963029&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85059963029&partnerID=8YFLogxK

U2 - 10.1002/jcp.28146

DO - 10.1002/jcp.28146

M3 - Article

C2 - 30637731

AN - SCOPUS:85059963029

VL - 234

SP - 15048

EP - 15060

JO - Journal of Cellular Physiology

JF - Journal of Cellular Physiology

SN - 0021-9541

IS - 9

ER -