Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-MYPT1 (Ser668) Antibody #3048

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

Applications Key:  W=Western Blotting
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

MYPT1 (Ser668) Antibody detects endogenous levels of MYPT1 only when phosphorylated at Ser668.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser668 of human MYPT1. Antibodies are purified using protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser668) Antibody (upper) or total MYPT1 Antibody #2634 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser668) Antibody. The phospho-specificity of the antibody was verified by peptide blocking using no peptide (left), phospho-Ser507 peptide (middle) or phospho-Ser668 peptide (right).

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).

Phospho-MYPT1 (Ser668) Antibody is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Ser668 was discovered using an Akt substrate antibody. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.

  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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