In vitro expression and functional analyses of the mutants p.R243Q, p.R241C and p.Y356X of the human phenylalanine hydroxylase

To study the expression of three phenylalanine hydroxylase (PAH) mutants (p.R243Q, p.R241C, and p.Y356X) and determine their pathogenicity. Bioinformatics techniques were used to predict the impact of PAH mutants on the structure and function of PAH protein. Corresponding mutant plasmids of PAH were...

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Published in:Zhongguo dang dai er ke za zhi 2024-02, Vol.26 (2), p.188-193
Main Authors: Pang, Yong-Hong, Gao, Xiang-Yu, Yuan, Zhen-Ya, Huang, Hui, Wang, Zeng-Qin, Peng, Lei, Li, Yi-Qun, Liu, Jie, Liu, Dong, Chen, Gui-Rong
Format: Article
Language:Chinese
Subjects:
Blotting, Western
Computational Biology
HEK293 Cells
Humans
Phenylalanine Hydroxylase - genetics
RNA, Messenger
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Summary:To study the expression of three phenylalanine hydroxylase (PAH) mutants (p.R243Q, p.R241C, and p.Y356X) and determine their pathogenicity. Bioinformatics techniques were used to predict the impact of PAH mutants on the structure and function of PAH protein. Corresponding mutant plasmids of PAH were constructed and expressed in HEK293T cells. Quantitative reverse transcription polymerase chain reaction was used to measure the mRNA expression levels of the three PAH mutants, and their protein levels were assessed using Western blot and enzyme-linked immunosorbent assay. Bioinformatics analysis predicted that all three mutants were pathogenic. The mRNA expression levels of the p.R243Q and p.R241C mutants in HEK293T cells were similar to the mRNA expression level of the wild-type control ( >0.05), while the mRNA expression level of the p.Y356X mutant significantly decreased (
ISSN:1008-8830
DOI:10.7499/j.issn.1008-8830.2309035