title: Quantitative assays for measuring human telomerase activity and DNA binding properties.
abstract: Telomerase is the ribonucleoprotein enzyme that catalyzes the processive addition of the telomeric DNA repeat 5'-TTAGGG-3' onto chromosome ends. In addition to its fascinating biochemical and enzymatic properties, clinical interest in telomerase stems from its dysregulated expression in ∼90% of human cancers, representing a broad spectrum of diseases. Exploiting telomerase as a therapeutic target and hence identifying and/or evaluating potential inhibitors requires quantitative measurement of its activity. This article presents procedures for measuring multiple aspects of telomerase enzymology that are relevant to both fundamental biochemistry and drug discovery: direct activity assays, DNA binding affinity, DNA dissociation, and cell-based over-expression of the active enzyme complex.
authors: Children's Medical Research Institute, 214 Hawkesbury Rd, Westmead, NSW 2145, Australia; University of Sydney, Sydney, NSW 2006, Australia.
authors: Christopher G
authors: CG
authors: Tomlinson
authors: Children's Medical Research Institute, 214 Hawkesbury Rd, Westmead, NSW 2145, Australia; University of Sydney, Sydney, NSW 2006, Australia.
authors: Natsuki
authors: N
authors: Sasaki
authors: Children's Medical Research Institute, 214 Hawkesbury Rd, Westmead, NSW 2145, Australia; University of Sydney, Sydney, NSW 2006, Australia.
authors: Julie
authors: J
authors: Jurczyluk
authors: Children's Medical Research Institute, 214 Hawkesbury Rd, Westmead, NSW 2145, Australia; University of Sydney, Sydney, NSW 2006, Australia.
authors: Tracy M
authors: TM
authors: Bryan
authors: Children's Medical Research Institute, 214 Hawkesbury Rd, Westmead, NSW 2145, Australia; University of Sydney, Sydney, NSW 2006, Australia. Electronic address: scohen@cmri.org.au.
authors: Scott B
authors: SB
authors: Cohen
mesh_terms: D002846:Chromatography, Affinity; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D019098:Telomerase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016615:Telomere / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004247:DNA; C509186:TERT protein, human; D019098:Telomerase
keywords: Activity assay; DNA affinity; Immunopurification; Telomerase