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Biophysical, Biochemical, and Cell Based Approaches Used to Decipher the Role of Carbonic Anhydrases in Cancer and to Evaluate the Potency of Targeted Inhibitors.
Mboge, Mam Y; Kota, Anusha; McKenna, Robert; Frost, Susan C.
Afiliación
  • Mboge MY; Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, P.O. Box 100245, Gainesville, FL 32610, USA.
  • Kota A; Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, P.O. Box 100245, Gainesville, FL 32610, USA.
  • McKenna R; Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, P.O. Box 100245, Gainesville, FL 32610, USA.
  • Frost SC; Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, P.O. Box 100245, Gainesville, FL 32610, USA.
Int J Med Chem ; 2018: 2906519, 2018.
Article en En | MEDLINE | ID: mdl-30112206
Carbonic anhydrases (CAs) are thought to be important for regulating pH in the tumor microenvironment. A few of the CA isoforms are upregulated in cancer cells, with only limited expression in normal cells. For these reasons, there is interest in developing inhibitors that target these tumor-associated CA isoforms, with increased efficacy but limited nonspecific cytotoxicity. Here we present some of the biophysical, biochemical, and cell based techniques and approaches that can be used to evaluate the potency of CA targeted inhibitors and decipher the role of CAs in tumorigenesis, cancer progression, and metastatic processes. These techniques include esterase activity assays, stop flow kinetics, and mass inlet mass spectroscopy (MIMS), all of which measure enzymatic activity of purified protein, in the presence or absence of inhibitors. Also discussed is the application of X-ray crystallography and Cryo-EM as well as other structure-based techniques and thermal shift assays to the studies of CA structure and function. Further, large-scale genomic and proteomic analytical methods, as well as cell based techniques like those that measure cell growth, apoptosis, clonogenicity, and cell migration and invasion, are discussed. We conclude by reviewing approaches that test the metastatic potential of CAs and how the aforementioned techniques have contributed to the field of CA cancer research.

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Int J Med Chem Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Int J Med Chem Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos