Molecular characterization of Thai patients with phenylalanine hydroxylase deficiency and in vitro functional study of two novel PAH variants.

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Tytuł:: Molecular characterization of Thai patients with phenylalanine hydroxylase deficiency and in vitro functional study of two novel PAH variants.
Autorzy:: Ngiwsara L; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.
Vatanavicharn N; Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. nithiwat_.
Sawangareetrakul P; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.
Liammongkolkul S; Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Ratanarak P; Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Boonyawat B; Department of Pediatrics, Phramongkutklao Hospital, Bangkok, Thailand.
Srisomsap C; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.
Champattanachai V; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.
Ketudat-Cairns J; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.; School of Biochemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
Wasant P; Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Svasti J; Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand.
Źródło:: Molecular biology reports [Mol Biol Rep] 2021 Mar; Vol. 48 (3), pp. 2063-2070. Date of Electronic Publication: 2021 Mar 07.
Typ publikacji:: Journal Article
Język:: English
Imprint Name(s):: Original Publication: Dordrecht, Boston, Reidel.
MeSH Terms:: Genetic Variation*
Phenylalanine Hydroxylase/*genetics
Phenylketonurias/*enzymology
Phenylketonurias/*genetics
Animals ; COS Cells ; Chlorocebus aethiops ; Gene Expression Regulation, Enzymologic ; Humans ; Mutation/genetics ; Phenotype ; Phenylalanine Hydroxylase/chemistry ; Thailand
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Grant Information:: BC-2020-05 Chulabhorn Research Institute; Siriraj Chalermprakiat Fund Faculty of Medicine Siriraj Hospital, Mahidol University
Contributed Indexing:: Keywords: PAH variants; Phenylalanine hydroxylase; Phenylketonuria; in vitro expression
Substance Nomenclature:: EC 1.14.16.1 (Phenylalanine Hydroxylase)
Entry Date(s):: Date Created: 20210307 Date Completed: 20210519 Latest Revision: 20210519
Update Code:: 20240104
DOI:: 10.1007/s11033-021-06163-w
PMID:: 33677757
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Phenylketonuria (PKU) is an autosomal recessive amino acid metabolism disorder caused by variants in the gene encoding phenylalanine hydroxylase (PAH; EC1.14.16.1). This study aimed to assess the specific heterogeneity of PAH variants found in Thai population as well as evaluate enzyme activity and expression of novel variants. PAH gene from 13 patients was analyzed by PCR amplification and direct Sanger-sequencing of 13 exons of the coding region. The novel variants were transiently transfected in COS-7 cells for functional verification. Eleven different PAH variants were identified: all pathogenic variants were missense variants, of which the most frequent variant was p.R169L, accounting for 24% (6/25) of all identified alleles. Two novel variants p.R169L and p.Y317N and previously reported variants with mutated residues at the same positions (p.R169H and p.Y317H) were expressed in COS-7 cells. These showed mildly impaired residual activity levels (42.3-63.1% of wild type), while the protein levels were well expressed (82.8-110%), except for p.R169L, which showed decreased protein expression of 55.7% compared to the wild type enzyme. All subjects with p.R169L identified in at least one of pathogenic alleles (one case is homozygous) had a metabolic phenotype of mild hyperphenylalaninemia (HPA). Our data has expanded the information on the genetic heterogeneity of Thai patients with PAH deficiency. This finding emphasizes the importance of genotyping in patients with HPA, and in vitro studies can provide additional information for prediction of phenotype.

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