PKCε regulates contraction-stimulated GLUT4 traffic in skeletal muscle cells.
The signaling pathways that stimulate glucose uptake in response to muscle contraction are not well defined. Recently, we showed that carbachol, an acetylcholine analog, stimulates contraction of C2C12 myotube cultures and the rapid arrival of myc-epitope tagged GLUT4 glucose transporters at the cell surface. Here, we explore a role for protein kinase C (PKC) in regulating GLUT4 traffic. Cell surface carbachol-induced GLUT4myc levels were partly inhibited by the conventional/novel PKC inhibitors GF-109203X, Gö6983, and Ro-31-8425 but not by the conventional PKC inhibitor Gö6976. C2C12 myotubes expressed several novel isoforms of PKC mRNA with PKCδ and PKCε in greater abundance. Carbachol stimulated phosphorylation of PKC isoforms and translocation of PKCδ and PKCε to membranes within 5 min. However, only a peptidic inhibitor of PKCε translocation (myristoylated-EAVSLKPT), but not one of PKCδ (myristoylated-SFNSYELGSL), prevented the GLUT4myc response to carbachol. Significant participation of PKCε in the carbachol-induced gain of GLUT4myc at the surface of C2C12 myotubes was further supported through siRNA-mediated PKCε protein knockdown. These findings support a role for novel PKC isoforms, especially PKCε, in contraction-stimulated GLUT4 traffic in muscle cells.
Niu
Wenyan
W
Department of Immunology, Tianjin Key Laboratory of Cellular and Molecular Immunology, Key Laboratory of Immuno Microenvironment and Disease of the Educational Ministry of China, Tianjin Medical University, Tianjin, China. wniu@tijmu.edu.cn
Bilan
Philip J
PJ
Yu
Junna
J
Gao
Jing
J
Boguslavsky
Shlomit
S
Schertzer
Jonathan D
JD
Chu
Guilan
G
Yao
Zhi
Z
Klip
Amira
A
D000263:Adenylate Kinase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002217:Carbachol / Q000008:administration & dosage / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000502:physiology*; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009119:Muscle Contraction / Q000502:physiology*; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism*; D051744:Protein Kinase C-epsilon / Q000235:genetics / Q000378:metabolism*
D051275:Glucose Transporter Type 4; C495693:Slc2a4 protein, mouse; D002217:Carbachol; D051744:Protein Kinase C-epsilon; D000263:Adenylate Kinase