Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb #4494

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC F H (M) (R) (Mk) Endogenous 78-82 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
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-DRP1 (Ser616) (D9A1) Rabbit mAb recognizes endogenous levels of DRP1 protein only when phosphorylated at Ser616.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser616 of human DRP1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or treated (+) with nocodazole (100 ng/ml, 17 hr), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb (upper) and DRP1 (D6C7) Rabbit mAb #8570 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (left) or λ phosphatase-treated (right), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb and propidium iodide (DNA content).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or λ phosphatase-treated (right), using Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

Dynamin-related protein 1 (DRP1) is a member of the dynamin superfamily of GTPases. Members of this family have diverse cellular functions including vesicle scission, organelle fission, viral resistance, and intracellular trafficking (reviewed in 1). DRP1 affects mitochondrial morphology and is important in mitochondrial and peroxisomal fission in mammalian cells (2-5). The yeast ortholog of DRP1 clusters into a spiral-shaped structure on the mitochondrial membrane at the site of fission (reviewed in 6), and this structure is likely conserved in mammalian cells (3). The division of the mitochondria, which is required for apoptosis, as well as normal cell growth and development is controlled, in part, by the phosphorylation of DRP1 at Ser616 by Cdk1/cyclin B and at Ser637 by protein kinase A (PKA) (reviewed in 6). When phosphorylated at Ser616, DRP1 stimulates mitochondrial fission during mitosis. Conversely, fission is inhibited when DRP1 is phosphorylated at Ser637 (reviewed in 6). Dephosphorylation at Ser637 by calcineurin reverses this inhibition (7). In addition to phosphorylation, sumoylation of DRP1 is also an enhancer of mitochondrial fission (8). Balancing fission and fusion events is essential for proper mitochondrial function. Research studies have demonstrated mitochondrial defects in a variety of neurodegenerative diseases including Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease (reviewed in 6).

  1. Praefcke, G.J. and McMahon, H.T. (2004) Nat Rev Mol Cell Biol 5, 133-47.
  2. Taguchi, N. et al. (2007) J Biol Chem 282, 11521-9.
  3. Smirnova, E. et al. (2001) Mol Biol Cell 12, 2245-56.
  4. Smirnova, E. et al. (1998) J Cell Biol 143, 351-8.
  5. Koch, A. et al. (2003) J Biol Chem 278, 8597-605.
  6. Knott, A.B. et al. (2008) Nat Rev Neurosci 9, 505-18.
  7. Cereghetti, G.M. et al. (2008) Proc Natl Acad Sci USA 105, 15803-8.
  8. Zunino, R. et al. (2007) J Cell Sci 120, 1178-88.

Application References

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

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