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Biochem Pharmacol ; 83(7): 969-76, 2012 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-22274639

RESUMO

Thiopurine S-methyltransferase (TPMT; EC 2.1.1.67) plays a pivotal role in thiopurine treatment outcomes. However, little has been known about its intracellular regulation. Here, we describe the effect of fluctuations in physiological levels of S-adenosyl-L-methionine (SAM) and related metabolites on TPMT activity levels in cell lines and erythrocytes from healthy donors. We determined higher TPMT activity in wild-type TPMT*1/*1 individuals with high SAM concentrations (n=96) compared to the low SAM level group (n=19; P<0.001). These findings confirm the results of our in vitro studies, which demonstrated that the restriction of L-methionine (Met) in cell growth media reversibly decreased TPMT activity and protein levels. Selective inhibition of distinct components of Met metabolism was used to demonstrate that SAM is implicitly responsible for direct post-translational TPMT stabilization. The greatest effect of SAM-mediated TPMT stabilization was observed in the case of wild-type TPMT*1 and variant *3C allozymes. In addition to TPMT genotyping, SAM may serve as an important biochemical marker in individualization of thiopurine therapy.


Assuntos
Metiltransferases/metabolismo , Processamento de Proteína Pós-Traducional , S-Adenosilmetionina/metabolismo , Técnicas de Cultura de Células , Meios de Cultura , Inibidores Enzimáticos/farmacologia , Eritrócitos/enzimologia , Ácido Fólico/metabolismo , Ácido Fólico/farmacologia , Genótipo , Células HEK293 , Células Hep G2 , Humanos , Isoenzimas , Metionina/metabolismo , Metionina/farmacologia , Metionina Adenosiltransferase/metabolismo , Metionina Adenosiltransferase/farmacologia , Metiltransferases/antagonistas & inibidores , Metiltransferases/genética , Estabilidade Proteica , S-Adenosilmetionina/antagonistas & inibidores , S-Adenosilmetionina/farmacologia
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