Publicação
Modulation of the Activity of Newly Synthesized Human Phenylalanine Hydroxylase Mutant Proteins by Low-Molecular-Weight Compounds
| Resumo: | Phenylketonuria, the most frequent disorder of amino acid metabolism, is caused by a deficient activity of human phenylalanine hydroxylase (hPAH). Rescue of the enzyme activity of several recombinant hPAH mutant forms (I65T, R261Q, R270K and V388M) by low molecular weight compounds namely glycerol, trimethylamine N-oxide (TMAO) and sodium 4-phenylbutyrate (4-PB) was investigated using a prokaryotic expression model. The studied compounds were added to the culture medium, in a concentration dependent manner, simultaneously to induction of protein expression. Among the tested molecules glycerol and TMAO were able to increase the enzyme activity of the studied mutant proteins. Furthermore, a decrease in aggregates and a recovery of the active tetrameric and dimeric forms were detected. Since the addition of the studied compounds to the medium did not change the expression level of E. Coli molecular chaperones we postulate that glycerol and TMAO rescue results from a direct stabilizing effect of the newly synthesized mutant hPAH enzymes.. - Fundacao para a Ciencia e Tecnologia, Portugal ; FEDER [MGI/POCTI/40844/2001, SFRH/BD/10807/2002, SFRH/BD/19024/2004]. - F. Ventura (iMed.UL) is gratefully acknowledged for critically reading of the manuscript. This work was supported by Fundacao para a Ciencia e Tecnologia, Portugal, and FEDER (MGI/POCTI/40844/2001, SFRH/BD/10807/2002 and SFRH/BD/19024/2004). |
|---|---|
| Autores principais: | Nascimento, Catia |
| Outros Autores: | Leandro, Joao; de Almeida, Isabel Tavares; Leandro, Paula |
| Assunto: | Biochemistry & Molecular Biology |
| Ano: | 2008 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso a metadados |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Phenylketonuria, the most frequent disorder of amino acid metabolism, is caused by a deficient activity of human phenylalanine hydroxylase (hPAH). Rescue of the enzyme activity of several recombinant hPAH mutant forms (I65T, R261Q, R270K and V388M) by low molecular weight compounds namely glycerol, trimethylamine N-oxide (TMAO) and sodium 4-phenylbutyrate (4-PB) was investigated using a prokaryotic expression model. The studied compounds were added to the culture medium, in a concentration dependent manner, simultaneously to induction of protein expression. Among the tested molecules glycerol and TMAO were able to increase the enzyme activity of the studied mutant proteins. Furthermore, a decrease in aggregates and a recovery of the active tetrameric and dimeric forms were detected. Since the addition of the studied compounds to the medium did not change the expression level of E. Coli molecular chaperones we postulate that glycerol and TMAO rescue results from a direct stabilizing effect of the newly synthesized mutant hPAH enzymes.. - Fundacao para a Ciencia e Tecnologia, Portugal ; FEDER [MGI/POCTI/40844/2001, SFRH/BD/10807/2002, SFRH/BD/19024/2004]. - F. Ventura (iMed.UL) is gratefully acknowledged for critically reading of the manuscript. This work was supported by Fundacao para a Ciencia e Tecnologia, Portugal, and FEDER (MGI/POCTI/40844/2001, SFRH/BD/10807/2002 and SFRH/BD/19024/2004). |
|---|
Atividades financiadas
Não foram encontrados projetos financiados.