Detection of APOBEC3 Proteins and Catalytic Activity in Urothelial Carcinoma

Methods Mol Biol. 2018:1655:97-107. doi: 10.1007/978-1-4939-7234-0_8.

Abstract

Members of the APOBEC3 (A3) family of enzymes were shown to act in an oncogenic manner in several cancer types. Immunodetection of APOBEC3A (A3A), APOBEC3B (A3B), and APOBEC3G (A3G) proteins is particularly challenging due to the large sequence homology of these proteins and limited availability of antibodies. Here we combine independent immunoblotting with an in vitro activity assay technique, to detect and categorize specific A3s expressed in urothelial bladder cancer and other cancer cells.

Keywords: APOBEC3; Cancer cell; Cytidine deaminase; Deamination assay; Mutation; Urothelial bladder cancer.

MeSH terms

  • APOBEC Deaminases
  • Catalysis
  • Cell Line
  • Cytidine Deaminase
  • Cytosine Deaminase / genetics
  • Cytosine Deaminase / metabolism*
  • DNA / metabolism
  • Enzyme Activation
  • Humans
  • Immunoblotting / methods
  • Multigene Family
  • Mutation
  • Urinary Bladder Neoplasms / diagnosis
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / metabolism
  • Urologic Neoplasms / diagnosis
  • Urologic Neoplasms / genetics
  • Urologic Neoplasms / metabolism*

Substances

  • DNA
  • Cytosine Deaminase
  • APOBEC Deaminases
  • APOBEC3 proteins, human
  • Cytidine Deaminase