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Identification and Validation of PTEN Complex, Associated Proteins

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Annual summary rept. 1 Nov 2004-31 Oct 2005

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The PTENMMAC1TEP1 is a tumor suppressor gene, targeted for biallelic somatic inactivation in a variety of cancers including advanced prostate adenocarcinomas among many others malignancies. PTEN is a phosphatase and has an important role in regulation of the PI3KAKT signaling pathway, which plays a key role in regulating cellular functions including proliferation, apoptosis, glucose homeostasis, cell size, nutrient response and DNA damage. Furthermore, PTEN functions in the cell to restrict both growth and survival in absence of growth signals. Studies performed in our laboratory indicated that in addition to its 47Kda form, PTEN could be detected as a part of a 600Kda complex. Further, we have also shown that PTEN acts as an antagonist of the PI3KAKT signaling, only when it is unphosphorylated and recruited into the large protein complex. To purify the complex and study the dynamics of the phosphorylated versus the unphosphorylated forms of PTEN in the cell we have generated a novel antiserum that selectively recognizes the unphosphorylated form of PTEN. This new antibody T382383 will allow us to study and purify the lower abundance of unphosphorylated form of PTEN present in the PTEN associated complex PAC. In addition, using our previously generated high affinity anti PTEN antibody C54 that recognizes both the phosphorylated and unphosphorylated forms of PTEN we generated an immunoaffinity column C54-DSSProtein A that maintains all the qualities of the C54 antibody, i.e. the high ability to bind and precipitate the endogenous PTEN. The immunoaffinity columns will be used additional step in our biochemical purification of the PAC.

Subject Categories:

  • Biochemistry
  • Anatomy and Physiology
  • Inorganic Chemistry

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