Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-MYPT1 (Thr853) Antibody #4563

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Hm Mk Dg Endogenous 140 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Dg=Dog
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-MYPT1 (Thr853) Antibody detects endogenous levels of MYPT1 only when phosphorylated at Thr853. The antibody cross-reacts with an unidentified protein at 40 kDa.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr853 of human MYPT1. Antibodies are purified using protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Phospho-MYPT1 (Thr853) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or treated with with the ROCK inhibitor Y27632 using Phospho-MYPT1 (Thr853) Antibody (upper) or MYPT1 Antibody #2634 (lower).

Background

Protein phosphatase 1 (PP1) is a ubiquitous eukaryotic protein serine/threonine phosphatase involved in the regulation of various cell functions. Substrate specificity is determined by the binding of a regulatory subunit to the PP1 catalytic subunit (PP1c). It is estimated that over fifty different regulatory subunits exist (1).The myosin phosphatase holoenzyme is composed of three subunits: PP1c, a targeting/regulatory subunit (MYPT/myosin-binding subunit of myosin phosphatase), and a 20 kDa subunit of unknown function (M20). MYPT binding to PP1cδ alters the conformation of the catalytic cleft and increases enzyme activity and specificity (2). Two MYPT isoforms that are 61% identical have been described. MYPT1 is widely expressed, while MYPT2 expression appears to be exclusive to heart and brain (3). Related family members include MBS85, MYPT3, and TIMAP (4).Myosin phosphatase regulates the interaction of actin and myosin in response to signaling through the small GTPase Rho. Rho activity inhibits myosin phosphatase via Rho-associated kinase (ROCK). Phosphorylation of MYPT1 at Thr696 and Thr853 results in phosphatase inhibition and cytoskeletal reorganization (5,6).

  1. Cohen, P.T. (2002) J Cell Sci 115, 241-56.
  2. Terrak, M. et al. (2004) Nature 429, 780-4.
  3. Fujioka, M. et al. (1998) Genomics 49, 59-68.
  4. Ito, M. et al. (2004) Mol Cell Biochem 259, 197-209.
  5. Birukova, A.A. et al. (2004) Microvasc Res 67, 64-77.
  6. Birukova, A.A. et al. (2004) J Cell Physiol 201, 55-70.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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