Analysis and biological relevance of advanced glycation end-products of DNA in eukaryotic cells

The FEBS Journal
Viola BreyerM Pischetsrieder

Abstract

Advanced glycation end-products (AGEs) of DNA are formed spontaneously by the reaction of carbonyl compounds such as sugars, methylglyoxal or dihydroxyacetone in vitro and in vivo. Little is known, however, about the biological consequences of DNA AGEs. In this study, a method was developed to determine the parameters that promote DNA glycation in cultured cells. For this purpose, the formation rate of N2-carboxyethyl-2'-deoxyguanosine (CEdG), a major DNA AGE, was measured in cultured hepatic stellate cells by liquid chromatography (LC)-MS/MS. In resting cells, a 1.7-fold increase of CEdG formation rate was observed during 14 days of incubation. To obtain insights into the functional consequences of DNA glycation, CEdG was introduced into a luciferase reporter gene vector and transfected into human embryonic kidney (HEK 293 T) cells. Gene activity was determined by chemiluminescence of the luciferase. Thus, CEdG adducts led to a dose-dependent and highly significant decrease in protein activity, which is caused by loss of functionality of the luciferase in addition to reduced transcription of the gene. When the CEdG-modified vector was transformed into Escherichia coli, a loss of ampicillin resistance was observed in comparison...Continue Reading

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Citations

Mar 5, 2013·Journal of Agricultural and Food Chemistry·Sabrina GensbergerMonika Pischetsrieder
Jul 11, 2012·International Journal of Cell Biology·Pamela Boon Li Pun, Michael P Murphy
May 8, 2014·Chemical Research in Toxicology·Katya V PetrovaCarmelo J Rizzo
Mar 23, 2011·Mutation Research·Georgia-Persephoni VoulgaridouAglaia Pappa
Feb 11, 2015·Analytical Chemistry·Goutam Kumar ChandraOlivier Piot
Jul 4, 2012·International Journal of Biological Macromolecules·Jalaluddin M AshrafKhursheed Alam
May 6, 2017·International Journal of Molecular Sciences·Sebastian BringsPeter P Nawroth
Dec 3, 2016·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Asif ZamanKhursheed Alam
Jul 20, 2017·Journal of Food Science and Technology·Chen WangLishuang Lv

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