Reevaluation of a DNA binding activity with specificity for chemically modified DNA
A DNA binding activity which appeared in direct filter binding assays to show specificity for DNA modified by N-acetoxy-2-acetylaminofluorene (AAF), N-methyl-N-nitrosourea and methylmethanesulfonate (Moranelli and Lieberman, Proc. Natl. Acad. Sci. USA 77, 3201, 1980) has been further purified and characterized. Equilibrium competition binding experiments indicate this activity shows little, or no, preferential binding of AAF-modified DNA compared to single-stranded DNA and only a small preference compared to u.v.-irradiated DNA or double-stranded DNA. These findings demonstrate the difficulty of using direct filter binding assays when purifying proteins with a desired biological function and the need to perform competition assays when evaluating specificity.
- Research Organization:
- Department of Pathology, Washington University School of Medicine, St. Louis, MO
- OSTI ID:
- 6579956
- Journal Information:
- Carcinogenesis (N.Y.); (United States), Journal Name: Carcinogenesis (N.Y.); (United States) Vol. 3:6; ISSN CRNGD
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560111* -- Radiation Effects on Biochemicals-- In Vitro-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AROMATICS
BIOASSAY
BIOCHEMICAL REACTION KINETICS
CARBONIC ACID DERIVATIVES
CHEMICAL BONDS
CONDENSED AROMATICS
DNA
ELECTROMAGNETIC RADIATION
ESTERS
FLUORENE
HYDROCARBONS
KINETICS
METHYL METHANESULFONATE
METHYL NITROSOUREA
MUTAGENS
NITROSO COMPOUNDS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
RADIATIONS
REACTION KINETICS
SPECIFICITY
SULFONIC ACID ESTERS
ULTRAVIOLET RADIATION