Subunit of mTORC1, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino acids. Growth factor-stimulated mTORC1 activation involves a AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-389', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Within mTORC1, AKT1S1 negatively regulates mTOR activity in a manner that is dependent on its phosphorylation state and binding to 14-3-3 proteins. Inhibits RHEB-GTP-dependent mTORC1 activation. Substrate for AKT1 phosphorylation, but can also be activated by AKT1-independent mechanisms. May also play a role in nerve growth factor-mediated neuroprotection.
Background References
1. Zhou Q et al. Targeting PRAS40: a novel therapeutic strategy for human diseases. J Drug Target. 2021 Aug
2. Qi Z et al. PRAS40 hyperexpression promotes hepatocarcinogenesis. EBioMedicine. 2020 Jan
Western blot analysis of Phospho-PRAS40 (T246) on different lysates with Rabbit anti-Phospho-PRAS40 (T246) antibody (HA721790) at 1/1,000 dilution.
Lane 1: HeLa cell lysate Lane 2: HeLa serum starved and treated with 100nM insulin for 5 minutes cell lysate Lane 3: HeLa serum starved and treated with 100nM insulin for 5 minutes, then treated with λpp for 1 hour cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 27 kDa Observed band size: 42 kDa
Exposure time: 2 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721790) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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