Involvement of calmodulin and myosin light chain kinase in activation of mTRPC5 expressed in HEK cells

Tae Kim Min, Joo Kim Byung, Hwa Lee Jae, Chun Kwon Seong, Soo Yeon Dong, Ki Yang Dong, Insuk So, Whan Kim Ki

Research output: Contribution to journalArticleResearchpeer-review

35 Citations (Scopus)

Abstract

The classic type of transient receptor potential channel (TRPC) is a molecular candidate for Ca2+-permeable cation channels in mammalian cells. Because TRPC channels have calmodulin (CaM) binding sites at their COOH termini, we investigated the effect of CaM on mTRPC5. TRPC5 was initially activated by muscarinic stimulation with 50 μM carbachol and then decayed rapidly even in the presence of carbachol. Intracellular CaM (150 μg/ml) increased the amplitude of mTRPC5 current activated by muscarinic stimulation. CaM antagonists (W-7 and calmidazolium) inhibited mTRPC5 currents when they were applied during the activation of mTRPC5. Pretreatment of W-7 and calmidazolium also inhibited the activation of mTRPC5 current. Inhibitors of myosin light chain kinase (MLCK) inhibited the activation of mTRPC5 currents, whereas inhibitors of CaM-dependent protein kinase II did not. Small interfering RNA against cardiac type MLCK also inhibited the activation of mTRPC5 currents. However, inhibitors of CaM or MLCK did not show any effect on GTPγS-induced currents. Application of both Rho kinase inhibitor and MLCK inhibitor inhibited GTPγS-induced currents. We conclude that CaM and MLCK modulates the activation process of mTRPC5.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume290
Issue number4
DOIs
StatePublished - 1 Apr 2006

Fingerprint

Myosin-Light-Chain Kinase
Calmodulin
calmidazolium
Transient Receptor Potential Channels
Carbachol
Cholinergic Agents
Cardiac Myosins
Calcium-Calmodulin-Dependent Protein Kinase Type 2
rho-Associated Kinases
Small Interfering RNA
Cations
Binding Sites

Keywords

  • Nonselective cation channel
  • Transient receptor potential channel

Cite this

Min, Tae Kim ; Byung, Joo Kim ; Jae, Hwa Lee ; Seong, Chun Kwon ; Dong, Soo Yeon ; Dong, Ki Yang ; So, Insuk ; Ki, Whan Kim. / Involvement of calmodulin and myosin light chain kinase in activation of mTRPC5 expressed in HEK cells. In: American Journal of Physiology - Cell Physiology. 2006 ; Vol. 290, No. 4.
@article{816b9027113a4266bba935104e35227b,
title = "Involvement of calmodulin and myosin light chain kinase in activation of mTRPC5 expressed in HEK cells",
abstract = "The classic type of transient receptor potential channel (TRPC) is a molecular candidate for Ca2+-permeable cation channels in mammalian cells. Because TRPC channels have calmodulin (CaM) binding sites at their COOH termini, we investigated the effect of CaM on mTRPC5. TRPC5 was initially activated by muscarinic stimulation with 50 μM carbachol and then decayed rapidly even in the presence of carbachol. Intracellular CaM (150 μg/ml) increased the amplitude of mTRPC5 current activated by muscarinic stimulation. CaM antagonists (W-7 and calmidazolium) inhibited mTRPC5 currents when they were applied during the activation of mTRPC5. Pretreatment of W-7 and calmidazolium also inhibited the activation of mTRPC5 current. Inhibitors of myosin light chain kinase (MLCK) inhibited the activation of mTRPC5 currents, whereas inhibitors of CaM-dependent protein kinase II did not. Small interfering RNA against cardiac type MLCK also inhibited the activation of mTRPC5 currents. However, inhibitors of CaM or MLCK did not show any effect on GTPγS-induced currents. Application of both Rho kinase inhibitor and MLCK inhibitor inhibited GTPγS-induced currents. We conclude that CaM and MLCK modulates the activation process of mTRPC5.",
keywords = "Nonselective cation channel, Transient receptor potential channel",
author = "Min, {Tae Kim} and Byung, {Joo Kim} and Jae, {Hwa Lee} and Seong, {Chun Kwon} and Dong, {Soo Yeon} and Dong, {Ki Yang} and Insuk So and Ki, {Whan Kim}",
year = "2006",
month = "4",
day = "1",
doi = "10.1152/ajpcell.00602.2004",
language = "English",
volume = "290",
journal = "American Journal of Physiology - Cell Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "4",

}

Involvement of calmodulin and myosin light chain kinase in activation of mTRPC5 expressed in HEK cells. / Min, Tae Kim; Byung, Joo Kim; Jae, Hwa Lee; Seong, Chun Kwon; Dong, Soo Yeon; Dong, Ki Yang; So, Insuk; Ki, Whan Kim.

In: American Journal of Physiology - Cell Physiology, Vol. 290, No. 4, 01.04.2006.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Involvement of calmodulin and myosin light chain kinase in activation of mTRPC5 expressed in HEK cells

AU - Min, Tae Kim

AU - Byung, Joo Kim

AU - Jae, Hwa Lee

AU - Seong, Chun Kwon

AU - Dong, Soo Yeon

AU - Dong, Ki Yang

AU - So, Insuk

AU - Ki, Whan Kim

PY - 2006/4/1

Y1 - 2006/4/1

N2 - The classic type of transient receptor potential channel (TRPC) is a molecular candidate for Ca2+-permeable cation channels in mammalian cells. Because TRPC channels have calmodulin (CaM) binding sites at their COOH termini, we investigated the effect of CaM on mTRPC5. TRPC5 was initially activated by muscarinic stimulation with 50 μM carbachol and then decayed rapidly even in the presence of carbachol. Intracellular CaM (150 μg/ml) increased the amplitude of mTRPC5 current activated by muscarinic stimulation. CaM antagonists (W-7 and calmidazolium) inhibited mTRPC5 currents when they were applied during the activation of mTRPC5. Pretreatment of W-7 and calmidazolium also inhibited the activation of mTRPC5 current. Inhibitors of myosin light chain kinase (MLCK) inhibited the activation of mTRPC5 currents, whereas inhibitors of CaM-dependent protein kinase II did not. Small interfering RNA against cardiac type MLCK also inhibited the activation of mTRPC5 currents. However, inhibitors of CaM or MLCK did not show any effect on GTPγS-induced currents. Application of both Rho kinase inhibitor and MLCK inhibitor inhibited GTPγS-induced currents. We conclude that CaM and MLCK modulates the activation process of mTRPC5.

AB - The classic type of transient receptor potential channel (TRPC) is a molecular candidate for Ca2+-permeable cation channels in mammalian cells. Because TRPC channels have calmodulin (CaM) binding sites at their COOH termini, we investigated the effect of CaM on mTRPC5. TRPC5 was initially activated by muscarinic stimulation with 50 μM carbachol and then decayed rapidly even in the presence of carbachol. Intracellular CaM (150 μg/ml) increased the amplitude of mTRPC5 current activated by muscarinic stimulation. CaM antagonists (W-7 and calmidazolium) inhibited mTRPC5 currents when they were applied during the activation of mTRPC5. Pretreatment of W-7 and calmidazolium also inhibited the activation of mTRPC5 current. Inhibitors of myosin light chain kinase (MLCK) inhibited the activation of mTRPC5 currents, whereas inhibitors of CaM-dependent protein kinase II did not. Small interfering RNA against cardiac type MLCK also inhibited the activation of mTRPC5 currents. However, inhibitors of CaM or MLCK did not show any effect on GTPγS-induced currents. Application of both Rho kinase inhibitor and MLCK inhibitor inhibited GTPγS-induced currents. We conclude that CaM and MLCK modulates the activation process of mTRPC5.

KW - Nonselective cation channel

KW - Transient receptor potential channel

UR - http://www.scopus.com/inward/record.url?scp=33646385634&partnerID=8YFLogxK

U2 - 10.1152/ajpcell.00602.2004

DO - 10.1152/ajpcell.00602.2004

M3 - Article

VL - 290

JO - American Journal of Physiology - Cell Physiology

JF - American Journal of Physiology - Cell Physiology

SN - 0363-6143

IS - 4

ER -