Acetyl CoA:arylamine N-acetyltransferase activity in rat hepatocytes cultured on different extracellular matrices

Toxicology in Vitro : an International Journal Published in Association with BIBRA
H ZaherC K Svensson

Abstract

N-Acetyltransferase (NAT) activity towards p-aminobenzoic acid and sulfamethazine was examined in primary cultures of rat hepatocytes cultured on three extracellular matrices (ECM)-type I collagen, thermally denatured type I collagen, and Matrigel((R)). Whereas protein and DNA content declined markedly during the first 24 hr of culture, p-acetylamidobenzoate (AcPABA) and N-acetylsulfamethazine (AcSMZ) formation were readily detectable on all three ECM for the 6-day culture period. Protein and DNA content, as well as NAT activities, were higher on Matrigel than on either of the other two ECM. Additional studies were conducted to confirm the expression of both enzymes during the culture period. The ratio of AcPABA to AcSMZ formation remained relatively stable throughout the 6-day culture period, suggesting that both enzymes continued to be expressed throughout the study period. Further studies in cells cultured on Matrigel revealed that AcPABA formation exhibited a time-dependent decline when cytosol from cultured cells was incubated at 50 degrees C, whereas AcSMZ formation proved to be thermostable. Moreover, methotrexate substantially inhibited AcPABA formation, but had only modest effects on AcSMZ. These studies support the co...Continue Reading

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Citations

Jun 2, 2015·Journal of Medicinal Chemistry·James J CrawfordJoachim Rudolph

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