The glutamate agonist homocysteine sulfinic acid stimulates glucose uptake through the calcium-dependent AMPK-p38 MAPK-protein kinase C ζ pathway in skeletal muscle cells

Ji Hae Kim, Jung Ok Lee, Soo Kyung Lee, Ji Wook Moon, Ga Young You, Su Jin Kim, Sun-Hwa Park, Ji Man Park, Se Young Lim, Pann Ghill Suh, Kyung Ok Uhm, Min Seok Song, Hyeon Soo Kim

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Abstract

Homocysteine sulfinic acid (HCSA) is a homologue of the amino acid cysteine and a selective metabotropic glutamate receptor (mGluR) agonist. However, the metabolic role of HCSA is poorly understood. In this study, we showed that HCSA and glutamate stimulated glucose uptake in C2C12 mouse myoblast cells and increased AMP-activated protein kinase (AMPK) phosphorylation. RT-PCR and Western blot analysis revealed that C2C12 expresses mGluR5. HCSA transiently increased the intracellular calcium concentration. Although α-methyl-4- carboxyphenylglycine, a metabotropic glutamate receptor antagonist, blocked the action of HCSA in intracellular calcium response and AMPK phosphorylation, 6-cyano-7-nitroquinoxaline-2,3-dione, an AMPA antagonist, did not exhibit such effects. Knockdown of mGluR5 with siRNA blocked HCSA-induced AMPK phosphorylation. Pretreatment of cells with STO-609, a calmodulin-dependent protein kinase kinase (CaMKK) inhibitor, blocked HCSA-induced AMPK phosphorylation, and knockdown of CaMKK blocked HCSA-induced AMPK phosphorylation. In addition, HCSA activated p38 mitogen-activated protein kinase (MAPK). Expression of dominant-negative AMPK suppressed HCSA-mediated phosphorylation of p38 MAPK, and inhibition of AMPK and p38 MAPK blocked HCSA-induced glucose uptake. Phosphorylation of protein kinase C ζ (PKCζ) was also increased by HCSA. Pharmacologic inhibition or knockdown of p38 MAPK blocked HCSA-induced PKCζ phosphorylation, and knockdown of PKCζ suppressed the HCSA-induced increase of cell surface GLUT4. The stimulatory effect of HCSA on cell surface GLUT4 was impaired in FITC-conjugated PKCζ siRNA-transfected cells. Together, the above results suggest that HCSA may have a beneficial role in glucose metabolism in skeletal muscle cells via stimulation of AMPK.

Original languageEnglish
Pages (from-to)7567-7576
Number of pages10
JournalJournal of Biological Chemistry
Volume286
Issue number9
DOIs
Publication statusPublished - 2011 Mar 4

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Excitatory Amino Acid Agonists
AMP-Activated Protein Kinases
Mitogen-Activated Protein Kinase Kinases
p38 Mitogen-Activated Protein Kinases
Muscle Cells
Protein Kinase C
Muscle
Skeletal Muscle
Cells
Calcium
Glucose
Phosphorylation
Calcium-Calmodulin-Dependent Protein Kinases
Metabotropic Glutamate Receptors
homocysteinesulfinic acid
Small Interfering RNA
Phosphotransferases
6-Cyano-7-nitroquinoxaline-2,3-dione
Excitatory Amino Acid Antagonists
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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The glutamate agonist homocysteine sulfinic acid stimulates glucose uptake through the calcium-dependent AMPK-p38 MAPK-protein kinase C ζ pathway in skeletal muscle cells. / Kim, Ji Hae; Lee, Jung Ok; Lee, Soo Kyung; Moon, Ji Wook; You, Ga Young; Kim, Su Jin; Park, Sun-Hwa; Park, Ji Man; Lim, Se Young; Suh, Pann Ghill; Uhm, Kyung Ok; Song, Min Seok; Kim, Hyeon Soo.

In: Journal of Biological Chemistry, Vol. 286, No. 9, 04.03.2011, p. 7567-7576.

Research output: Contribution to journalArticle

Kim, Ji Hae ; Lee, Jung Ok ; Lee, Soo Kyung ; Moon, Ji Wook ; You, Ga Young ; Kim, Su Jin ; Park, Sun-Hwa ; Park, Ji Man ; Lim, Se Young ; Suh, Pann Ghill ; Uhm, Kyung Ok ; Song, Min Seok ; Kim, Hyeon Soo. / The glutamate agonist homocysteine sulfinic acid stimulates glucose uptake through the calcium-dependent AMPK-p38 MAPK-protein kinase C ζ pathway in skeletal muscle cells. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 9. pp. 7567-7576.
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abstract = "Homocysteine sulfinic acid (HCSA) is a homologue of the amino acid cysteine and a selective metabotropic glutamate receptor (mGluR) agonist. However, the metabolic role of HCSA is poorly understood. In this study, we showed that HCSA and glutamate stimulated glucose uptake in C2C12 mouse myoblast cells and increased AMP-activated protein kinase (AMPK) phosphorylation. RT-PCR and Western blot analysis revealed that C2C12 expresses mGluR5. HCSA transiently increased the intracellular calcium concentration. Although α-methyl-4- carboxyphenylglycine, a metabotropic glutamate receptor antagonist, blocked the action of HCSA in intracellular calcium response and AMPK phosphorylation, 6-cyano-7-nitroquinoxaline-2,3-dione, an AMPA antagonist, did not exhibit such effects. Knockdown of mGluR5 with siRNA blocked HCSA-induced AMPK phosphorylation. Pretreatment of cells with STO-609, a calmodulin-dependent protein kinase kinase (CaMKK) inhibitor, blocked HCSA-induced AMPK phosphorylation, and knockdown of CaMKK blocked HCSA-induced AMPK phosphorylation. In addition, HCSA activated p38 mitogen-activated protein kinase (MAPK). Expression of dominant-negative AMPK suppressed HCSA-mediated phosphorylation of p38 MAPK, and inhibition of AMPK and p38 MAPK blocked HCSA-induced glucose uptake. Phosphorylation of protein kinase C ζ (PKCζ) was also increased by HCSA. Pharmacologic inhibition or knockdown of p38 MAPK blocked HCSA-induced PKCζ phosphorylation, and knockdown of PKCζ suppressed the HCSA-induced increase of cell surface GLUT4. The stimulatory effect of HCSA on cell surface GLUT4 was impaired in FITC-conjugated PKCζ siRNA-transfected cells. Together, the above results suggest that HCSA may have a beneficial role in glucose metabolism in skeletal muscle cells via stimulation of AMPK.",
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AU - Moon, Ji Wook

AU - You, Ga Young

AU - Kim, Su Jin

AU - Park, Sun-Hwa

AU - Park, Ji Man

AU - Lim, Se Young

AU - Suh, Pann Ghill

AU - Uhm, Kyung Ok

AU - Song, Min Seok

AU - Kim, Hyeon Soo

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