cdc25C Antibody Sampler Kit #9555
Product Information
Kit Usage Information
Protocols
Product Description
The cdc25C Antibody Sampler Kit provides an economical means to investigate the entry of eukaryotic cells into mitosis. The kit contains enough primary and secondary antibodies to perform two Western blots with each antibody.
Background
Cdc25 is a protein phosphatase responsible for dephosphorylating and activating cdc2, a crucial step in regulating the entry of all eukaryotic cells into mitosis (1). cdc25C is constitutively phosphorylated at Ser216 throughout interphase by c-TAK1, while phosphorylation at this site is DNA damage-dependent at the G2/M checkpoint (2). When phosphorylated at Ser216, cdc25C binds to members of the 14-3-3 family of proteins, sequestering cdc25C in the cytoplasm and thereby preventing premature mitosis (3). The checkpoint kinases Chk1 and Chk2 phosphorylate cdc25C at Ser216 in response to DNA damage (4,5).
Full activation of cdc25C involves phosphorylation at more than 12 different sites by cdc2/cyclin B and Polo-like kinase, and the activity of Pin1, a peptidyl-prolyl isomerase (PPI) (6,7). Pin1 contains a WW domain that binds phospho-Ser/Thr-Pro sites and a catalytic PPI region that induces a cis/trans isomerization on phospho-Ser/Thr-Pro bonds (8). Thr48 and Thr67 of cdc25C interact directly with the WW domain of Pin1 when these sites are phosphorylated (9). Thr48 phosphorylation also mediates binding to CKS/p13SUC1 (10).
Full activation of cdc25C involves phosphorylation at more than 12 different sites by cdc2/cyclin B and Polo-like kinase, and the activity of Pin1, a peptidyl-prolyl isomerase (PPI) (6,7). Pin1 contains a WW domain that binds phospho-Ser/Thr-Pro sites and a catalytic PPI region that induces a cis/trans isomerization on phospho-Ser/Thr-Pro bonds (8). Thr48 and Thr67 of cdc25C interact directly with the WW domain of Pin1 when these sites are phosphorylated (9). Thr48 phosphorylation also mediates binding to CKS/p13SUC1 (10).
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- Kumagai, A. and Dunphy, W.G. (1999) Genes Dev. 13, 1067-1072.
- Blasina, A. et al. (1999) Curr. Biol. 9, 1-10.
- Furnari, B. et al. (1999) Mol. Biol. Cell 10, 833-845.
- Izumi, T. and Maller, J.L. (1993) Mol Biol Cell 4, 1337-50.
- Stukenberg, P.T. and Kirschner, M.W. (2001) Mol Cell 7, 1071-83.
- Yaffe, M.B. et al. (1997) Science 278, 1957-60.
- Lu, P.J. et al. (1999) Science 283, 1325-8.
- Landrieu, I. et al. (2001) J Biol Chem 276, 1434-8.
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