Enzyme kinetic properties of human recombinant arylamine N-acetyltransferase 2 allotypic variants expressed in Escherichia coli

Biochemical Pharmacology
D HickmanE Sim

Abstract

Arylamine N-acetyltransferase (NAT2) catalyses the N-acetylation of primary arylamine and hydrazine drugs and chemicals. N-Acetylation is subject to polymorphism, and humans can be categorized as either fast or slow acetylators according to their ability to N-acetylate certain arylamine substrates in vivo. Genetic variants at the polymorphic NAT2 locus have been described. We expressed five of the most common NAT2 variants (NAT2 4, NAT2 5A, NAT2 5B, NAT2 6A and NAT2 7B) in Escherichia coli as a convenient source of the human variants. The apparent Km values (at 100 microM acetyl CoA as co-substrate) of the different NAT2 variants for sulphamethazine, dapsone, p-anisidine, 2-aminofluorene, procainamide and isoniazid were determined. Data show that the apparent Km of the slow variant NAT2 7B for the arylamine sulphamethazine was 10-fold lower than all the other allotypes. The apparent Km for the structurally related sulphone antibiotic dapsone was 5-fold lower for the slow variant NAT2 7B when compared with the wild-type NAT2 4. These results indicate that the NAT2 7B specific amino acid substitution, Gly286-Glu, is important in promoting the binding of sulphamethazine and dapsone to the active site.

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Nov 3, 1976·Journal of Chromatography·K D HaegeleJ L McNay
Dec 1, 1992·Pharmacogenetics·J R IdleH Hadidi
Jun 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·M BlumU A Meyer
Jul 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·K P VatsisW W Weber
Jun 1, 1974·The Biochemical Journal·R Eisenthal, A Cornish-Bowden
Feb 1, 1995·Pharmacogenetics·K P VatsisM V Relling

Citations

Oct 7, 2000·Bulletin of Experimental Biology and Medicine·S I MakarovaS M Gavalov
Oct 15, 2005·Bulletin of Experimental Biology and Medicine·S I MakarovaV V Lyakhovich
Nov 21, 2007·Bulletin of Experimental Biology and Medicine·M V NikishinaV V Lyakhovich
May 12, 1997·Mutation Research·D M GrantR Grewal
Dec 1, 1995·Trends in Genetics : TIG·E SimP Thygesen
Oct 20, 2011·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Lori M MillnerD W Hein
Nov 1, 2005·Drug Metabolism Reviews·S Boukouvala, G Fakis
Mar 26, 2015·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Azza M KamelHeba S Moussa
Oct 7, 2010·BMC Genetics·Jhimmy TalbotFabrício Rios-Santos
Oct 7, 2016·Acta Pharmaceutica Sinica. B·Pengcheng WangXiaochao Ma
Sep 11, 2016·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Ameeruddin Nusrath UnissaNagamiah Selvakumar
May 7, 2002·The Pharmacogenomics Journal·N J ButcherR F Minchin
Apr 19, 2015·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Nathalie RiouxNigel J Waters
Jul 28, 2017·Tuberculosis and Respiratory Diseases·Ameeruddin Nusrath UnissaLuke Elizabeth Hanna
Jun 20, 2017·Pharmacogenomics·D W Hein, M A Doll
Nov 27, 1999·The Journal of Biological Chemistry·M A LeffD W Hein

Related Concepts

NAT2 protein, human
Arylamine N-Acetyltransferase
DNA Sequence
DNA, Double-Stranded
Alkalescens-Dispar Group
Shuttle Vectors
Alloenzymes
Proteins, Recombinant DNA

Related Feeds

Antitubercular Agents

Antitubercular agents are pharmacologic agents for treatment of tuberculosis. Discover the latest research on antitubercular agents here.

Antitubercular Agents (ASM)

Antitubercular agents are pharmacologic agents for treatment of tuberculosis. Discover the latest research on antitubercular agents here.