Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-Tuberin/TSC2 (Ser1387) Antibody #5584

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk Endogenous 200 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Tuberin/TSC2 (Ser1387) Antibody detects endogenous levels of tuberin protein only when phosphorylated at Ser1387. This antibody also cross-reacts with an unidentified 140 kD protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser1387 of human tuberin protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with AICAR #9944 (2 mM for 30 minutes), using Phospho-Tuberin/TSC2 (Ser1387) Antibody (upper) and Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, treated with AICAR #9944 (2 mM for 30 minutes), using Phospho-Tuberin/TSC2 (Ser1387) Antibody. The phospho-specificity of the antibody was verified by treating the membrane with (+) or without (-) calf intestinal phosphatase (CIP) after western transfer.

Background

Tuberin is a product of the TSC2 tumor suppressor gene and an important regulator of cell proliferation and tumor development (1). Mutations in either TSC2 or the related TSC1 (hamartin) gene cause tuberous sclerosis complex (TSC), an autosomal dominant disorder characterized by development of multiple, widespread non-malignant tumors (2). Tuberin is directly phosphorylated at Thr1462 by Akt/PKB (3). Phosphorylation at Thr1462 and Tyr1571 regulates tuberin-hamartin complexes and tuberin activity (3-5). In addition, tuberin inhibits the mammalian target of rapamycin (mTOR), which promotes inhibition of p70 S6 kinase, activation of eukaryotic initiation factor 4E binding protein 1 (4E-BP1, an inhibitor of translation initiation), and eventual inhibition of translation (3,6,7).

Phosphorylation of tuberin by AMPK at Ser1387 is necessary for cell size control in response to energy deprivation and protects from apoptosis (8). Furthermore, phosphorylation at Ser1387 primes phosphorylation by GSK3 of upstream sites (Ser1383, Ser1379 and Ser1375), integrating Wnt signaling (9).

  1. Soucek, T. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 15653-15658.
  2. Sparagana, S.P. and Roach, E.S. (2000) Curr. Opin. Neurol. 13, 115-119.
  3. Manning, B. D. et al. (2002) Mol. Cell 10, 151-161.
  4. Aicher, L. D. et al. (2001) J. Biol. Chem. 276, 21017-21021.
  5. Dan, H. C. et al. (2002) J. Biol. Chem. 277, 35364-35370.
  6. Goncharova, E.A. et al. (2002) J. Biol. Chem. 277, 30958-30967.
  7. Inoki, K. et al. (2002) Nat. Cell Biol. 4, 648-657.
  8. Inoki, K. et al. (2003) Cell 115, 577-590.
  9. Inoki, K. et al. (2006) Cell 126, 955-968.

Application References

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