Identification of a novel phosphorylation site on TBC1D4 regulated by AMP-activated protein kinase in skeletal muscle
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Identification of a novel phosphorylation site on TBC1D4 regulated by AMP-activated protein kinase in skeletal muscle. / Treebak, Jonas Thue; Taylor, Eric B.; Witczak, Carol A.; An, Ding; Toyoda, Taro; Koh, Ho-Jin; Xie, Jianxin; Feener, Edward P.; Wojtaszewski, Jørgen; Hirshman, Michael F.; Goodyear, Laurie J.
In: American Journal of Physiology: Cell Physiology, Vol. 298, No. 2, 2010, p. C377-C385.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Identification of a novel phosphorylation site on TBC1D4 regulated by AMP-activated protein kinase in skeletal muscle
AU - Treebak, Jonas Thue
AU - Taylor, Eric B.
AU - Witczak, Carol A.
AU - An, Ding
AU - Toyoda, Taro
AU - Koh, Ho-Jin
AU - Xie, Jianxin
AU - Feener, Edward P.
AU - Wojtaszewski, Jørgen
AU - Hirshman, Michael F.
AU - Goodyear, Laurie J.
N1 - CURIS 2010 5200 013
PY - 2010
Y1 - 2010
N2 - TBC1D4 (also known as AS160) regulates GLUT4 translocation and glucose uptake in adipocytes and skeletal muscle. Its mode of action involves phosphorylation of Serine (S)/Threonine (T) residues by upstream kinases resulting in inactivation of Rab-GAP activity leading to GLUT4 mobilization. The majority of known phosphorylation sites on TBC1D4 lie within the Akt consensus motif and are phosphorylated by insulin stimulation. However, the 5 AMP activated protein kinase (AMPK) and other kinases may also phosphorylate TBC1D4, and therefore we hypothesized the presence of additional phosphorylation sites. Mouse skeletal muscles were contracted or stimulated with 5-aminoimidazole-4-carboxmide riboside (AICAR) and muscle lysates were subjected to mass spectrometry analyses resulting in identification of novel putative phosphorylation sites on TBC1D4. The surrounding amino acid sequence predicted that S711 would be recognized by AMPK. Using a phospho-specific antibody against S711, we found that AICAR and contraction increased S711 phosphorylation in mouse skeletal muscle and this increase was abolished in muscle-specific AMPKalpha2 kinase dead transgenic mice. Exercise in human vastus lateralis muscle also increased TBC1D4 S711 phosphorylation. Recombinant AMPK, but not Akt1, Akt2, or PKCzeta phosphorylated purified muscle TBC1D4 on S711 in vitro. Interestingly, S711 was also phosphorylated in response to insulin in an Akt2- and rapamycin-independent, but a wortmannin-sensitive manner, suggesting this site is regulated by one or more additional upstream kinases. Despite increased S711 phosphorylation with AICAR, contraction, and insulin, mutation of S711 to alanine did not alter glucose uptake in response to these stimuli. S711 is a novel TBC1D4 phosphorylation site regulated by AMPK in skeletal muscle.
AB - TBC1D4 (also known as AS160) regulates GLUT4 translocation and glucose uptake in adipocytes and skeletal muscle. Its mode of action involves phosphorylation of Serine (S)/Threonine (T) residues by upstream kinases resulting in inactivation of Rab-GAP activity leading to GLUT4 mobilization. The majority of known phosphorylation sites on TBC1D4 lie within the Akt consensus motif and are phosphorylated by insulin stimulation. However, the 5 AMP activated protein kinase (AMPK) and other kinases may also phosphorylate TBC1D4, and therefore we hypothesized the presence of additional phosphorylation sites. Mouse skeletal muscles were contracted or stimulated with 5-aminoimidazole-4-carboxmide riboside (AICAR) and muscle lysates were subjected to mass spectrometry analyses resulting in identification of novel putative phosphorylation sites on TBC1D4. The surrounding amino acid sequence predicted that S711 would be recognized by AMPK. Using a phospho-specific antibody against S711, we found that AICAR and contraction increased S711 phosphorylation in mouse skeletal muscle and this increase was abolished in muscle-specific AMPKalpha2 kinase dead transgenic mice. Exercise in human vastus lateralis muscle also increased TBC1D4 S711 phosphorylation. Recombinant AMPK, but not Akt1, Akt2, or PKCzeta phosphorylated purified muscle TBC1D4 on S711 in vitro. Interestingly, S711 was also phosphorylated in response to insulin in an Akt2- and rapamycin-independent, but a wortmannin-sensitive manner, suggesting this site is regulated by one or more additional upstream kinases. Despite increased S711 phosphorylation with AICAR, contraction, and insulin, mutation of S711 to alanine did not alter glucose uptake in response to these stimuli. S711 is a novel TBC1D4 phosphorylation site regulated by AMPK in skeletal muscle.
U2 - 10.1152/ajpcell.00297.2009
DO - 10.1152/ajpcell.00297.2009
M3 - Journal article
C2 - 19923418
VL - 298
SP - C377-C385
JO - American Journal of Physiology: Cell Physiology
JF - American Journal of Physiology: Cell Physiology
SN - 0363-6143
IS - 2
ER -
ID: 17112069