Definitive characterization of human thymine glycol N-glycosylase activity

Journal Article · Tue Mar 24 04:00:00 EST 1987 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00380a029· OSTI ID:6470481

Higgins, S A; Frenkel, K; Cummings, A; Teebor, G W

An N-glycosylase activity that released cis-(/sup 3/H)-5,6-dihydroxy-5,6-dihydrothymine (thymine glycol, TG) from chemically oxidized poly(dA-(/sup 3/H)dT) was unambiguously characterized both in extracts of HeLa cells and in purified Escherichia coli endonuclease III. This was accomplished by use of a microderivatization procedure that quantitatively converted cis-TG to 5-hydroxy-5-methylhydantoin (HMH). The reaction products were analyzed by high-pressure liquid chromatography before and after derivation by using cis-(/sup 14/C)TG and (/sup 14/C)HMH, which had been independently synthesized, as reference compounds. This technique facilitated construction of a v/(E)/sub t/ plot for the enzyme activity in HeLa cells, permitting estimation of its specific activity. The results obtained prove the existence of both human and bacterial N-glycosylase activities that effect removal of TG from DNA.

• Research Organization: New York Univ. Medical Center, NY
• OSTI ID: 6470481
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:6; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALCOHOLS
ANIMALS
AZINES
BACTERIA
BIOCHEMISTRY
CARBON 14 COMPOUNDS
CHEMICAL REACTIONS
CHEMISTRY
CHROMATOGRAPHY
DERIVATIZATION
DNA
ENZYME ACTIVITY
ENZYMES
ESCHERICHIA COLI
ESTERASES
GLYCOLS
GLYCOSYL HYDROLASES
HELA CELLS
HETEROCYCLIC COMPOUNDS
HYDROLASES
HYDROXY COMPOUNDS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MAN
MICROORGANISMS
NUCLEASES
NUCLEIC ACIDS
O-GLYCOSYL HYDROLASES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXIDATION
PHOSPHODIESTERASES
PRIMATES
PYRIMIDINES
SEPARATION PROCESSES
THYMINE
TRITIUM COMPOUNDS
URACILS
VERTEBRATES