Regulatory mechanisms of SNAT2, an amino acid transporter, in L6 rat skeletal muscle cells by insulin, osmotic shock and amino acid deprivation

被引:23
|
作者
Kashiwagi, Hitoshi [1 ]
Yamazaki, Kojiro [1 ]
Takekuma, Yoh [1 ]
Ganapathy, Vadivel [2 ]
Sugawara, Mitsuru [1 ]
机构
[1] Hokkaido Univ Hosp, Dept Pharm, Sapporo, Hokkaido 060, Japan
[2] Med Coll Georgia, Dept Biochem & Mol Biol, Augusta, GA 30912 USA
关键词
SNAT2; Insulin stimulation; Osmotic shock; Amino acid deprivation; Transporter recruitment; de novo protein synthesis; TISSUE EXPRESSION PATTERN; CANINE KIDNEY-CELLS; SYSTEM-A; FUNCTIONAL-CHARACTERISTICS; ADAPTIVE REGULATION; PLASMA-MEMBRANE; GENE; ATA2; ELEMENT; PROTEIN;
D O I
10.1007/s00726-008-0050-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Several studies have demonstrated that the activity of system A is upregulated by insulin, osmotic shock and amino acid deprivation. However, the mechanisms are not clear. We carried out studies using L6 rat skeletal muscle cells to clarify the mechanisms of upregulation of system A activity by insulin, osmotic shock and amino acid deprivation. The upregulation was found to be due to an increase in V (max), not K (m). Chloroquine and wortmannin inhibited the upregulation induced by insulin stimulation and amino acid deprivation but not that induced by osmotic shock. On the other hand, cycloheximide and actinomycin D inhibited the upregulation by each stimulation. Moreover, PD98059 and SP600125 inhibited only amino acid deprivation-induced upregulation and SB202190 inhibited only insulin-induced upregulation. Our findings indicate that the mechanisms of upregulation of system A activity by insulin, osmotic shock and amino acid deprivation are different in L6 cells. Western blot and RT-PCR analysis showed an increase in system A at the protein and mRNA levels with each stimulation.
引用
收藏
页码:219 / 230
页数:12
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