| Arginine supplementation required for adequate nutrient requirement to stimulate anabolic processes, such as protein synthesis and proliferation. The purpose of this study is to discover the effect supplementation of arginine on identified protein that functions in pathway of protein synthesis in HepG2cells. HepG2cells and arginine were used in this study as the main materials. The main methods that were used are cell culture, iTRAQ and western blot. The software packages were utilized for data analysis:Mascot for identification and quantification of differentially expressed proteins and String to provide insight into COG function. The result of arginine supplementation via iTRAQ assay showed that total differentially expressed protein on resupplementation group is higher than deprivation group and most of protein that detected is up-regulated protein. On deprivation group found decreased activity of AMPK on mTOR pathway, AMPK and Atg8activity in regulation of autophagy pathway. Decreasing AMPK will increase mTOR activity that will inhibit autophagy and induce protein synthesis in HepG2cells. For GO analysis can be known that the identified proteins represent the relevance and variety of molecular functions on binding and catalytic activity, acts in cellular and metabolic process, and large number found located in cell and organelle on all groups. The important protein that find in this study is pyroline-5-carboxylate dehydrogenase converts glutamyl-y-semialdehyde into glutamate. Glutamate is the result of arginine conversion, which has role in protein formation. Validation by Western Blot analysis shown during8h arginine deprivation decreases level of p62and LC3, but increase S6K and phosphorylation of mTOR by4mM arginine resupplementation in HepG2cells. In conclusion, arginine resupplementation will increase identified protein, inhibit autophagy, and can used for protein synthesis via increasing mTOR activity in HepG2cells. |
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