Structural and Functional Impact of Seven Missense Variants of
Phenylalanine Hydroxylase

J 2019

Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase

PECIMONOVA, Martina, Daniela KLUCKOVA, František CSICSAY, Kamila RÉBLOVÁ, Jan KRAHULEC et. al.

Basic information

Original name

Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase

Name in Czech

Strukturální a funkční vliv sedmi missense patogennich sekvencnich variant na enzym fenylalanin hydroxylazu

Authors

PECIMONOVA, Martina (703 Slovakia), Daniela KLUCKOVA (703 Slovakia), František CSICSAY (703 Slovakia), Kamila RÉBLOVÁ (203 Czech Republic, belonging to the institution), Jan KRAHULEC (703 Slovakia), Dagmar PROCHÁZKOVÁ (203 Czech Republic, belonging to the institution), Ludovit ŠKULTETY (703 Slovakia), Ludevít KADASI (703 Slovakia) and Andrea ŠOLTYSOVÁ (703 Slovakia, guarantor)

Edition

Genes, Basilej, MDPI, 2019, 2073-4425

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10603 Genetics and heredity

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.759

RIV identification code

RIV/00216224:14110/19:00109997

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.3390/genes10060459

UT WoS

000473797000054

Keywords (in Czech)

fenylalnin hydroxylaza fenylketonurie BH4 funkční studie missense varianty

Keywords in English

Phenylalanine hydroxylase phenylketonuria BH4 functional studies missense variants

Abstract

V originále

The molecular genetics of well-characterized inherited diseases, such as phenylketonuria (PKU) and hyperphenylalaninemia (HPA) predominantly caused by mutations in the phenylalanine hydroxylase (PAH) gene, is often complicated by the identification of many novel variants, often with no obvious impact on the associated disorder. To date, more than 1100 PAH variants have been identified of which a substantial portion have unknown clinical significance. In this work, we study the functionality of seven yet uncharacterized PAH missense variants p.Asn167Tyr, p.Thr200Asn, p.Asp229Gly, p.Gly239Ala, p.Phe263Ser, p.Ala342Pro, and p.Ile406Met first identified in the Czech PKU/HPA patients. From all tested variants, three of them, namely p.Asn167Tyr, p.Thr200Asn, and p.Ile406Met, exerted residual enzymatic activity in vitro similar to wild type (WT) PAH, however, when expressed in HepG2 cells, their protein level reached a maximum of 72.1% ± 4.9%, 11.2% ± 4.2%, and 36.6% ± 7.3% compared to WT PAH, respectively. Remaining variants were null with no enzyme activity and decreased protein levels in HepG2 cells. The chaperone-like effect of applied BH4 precursor increased protein level significantly for p.Asn167Tyr, p.Asp229Gly, p.Ala342Pro, and p.Ile406Met. Taken together, our results of functional characterization in combination with in silico prediction suggest that while p.Asn167Tyr, p.Thr200Asn, and p.Ile406Met PAH variants have a mild impact on the protein, p.Asp229Gly, p.Gly239Ala, p.Phe263Ser, and p.Ala342Pro severely affect protein structure and function.

Citovat

PECIMONOVA, Martina, Daniela KLUCKOVA, František CSICSAY, Kamila RÉBLOVÁ, Jan KRAHULEC, Dagmar PROCHÁZKOVÁ, Ludovit ŠKULTETY, Ludevít KADASI and Andrea ŠOLTYSOVÁ. Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase. Genes. Basilej: MDPI, 2019, vol. 10, No 6, p. 1-13. ISSN 2073-4425. Available from: https://dx.doi.org/10.3390/genes10060459.

@article{1542176,
   author = {Pecimonova, Martina and Kluckova, Daniela and Csicsay, František and Réblová, Kamila and Krahulec, Jan and Procházková, Dagmar and Škultety, Ludovit and Kadasi, Ludevít and Šoltysová, Andrea},
   article_location = {Basilej},
   article_number = {6},
   doi = {http://dx.doi.org/10.3390/genes10060459},
   keywords = {Phenylalanine hydroxylase phenylketonuria BH4 functional studies missense variants},
   language = {eng},
   issn = {2073-4425},
   journal = {Genes},
   title = {Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase},
   url = {https://www.mdpi.com/2073-4425/10/6/459},
   volume = {10},
   year = {2019}
}

TY  - JOUR
ID  - 1542176
AU  - Pecimonova, Martina - Kluckova, Daniela - Csicsay, František - Réblová, Kamila - Krahulec, Jan - Procházková, Dagmar - Škultety, Ludovit - Kadasi, Ludevít - Šoltysová, Andrea
PY  - 2019
TI  - Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase
JF  - Genes
VL  - 10
IS  - 6
SP  - 1-13
EP  - 1-13
PB  - MDPI
SN  - 20734425
KW  - Phenylalanine hydroxylase phenylketonuria BH4 functional studies missense variants
UR  - https://www.mdpi.com/2073-4425/10/6/459
L2  - https://www.mdpi.com/2073-4425/10/6/459
N2  - The molecular genetics of well-characterized inherited diseases, such as phenylketonuria (PKU) and hyperphenylalaninemia (HPA) predominantly caused by mutations in the phenylalanine hydroxylase (PAH) gene, is often complicated by the identification of many novel variants, often with no obvious impact on the associated disorder. To date, more than 1100 PAH variants have been identified of which a substantial portion have unknown clinical significance. In this work, we study the functionality of seven yet uncharacterized PAH missense variants p.Asn167Tyr, p.Thr200Asn, p.Asp229Gly, p.Gly239Ala, p.Phe263Ser, p.Ala342Pro, and p.Ile406Met first identified in the Czech PKU/HPA patients. From all tested variants, three of them, namely p.Asn167Tyr, p.Thr200Asn, and p.Ile406Met, exerted residual enzymatic activity in vitro similar to wild type (WT) PAH, however, when expressed in HepG2 cells, their protein level reached a maximum of 72.1% ± 4.9%, 11.2% ± 4.2%, and 36.6% ± 7.3% compared to WT PAH, respectively. Remaining variants were null with no enzyme activity and decreased protein levels in HepG2 cells. The chaperone-like effect of applied BH4 precursor increased protein level significantly for p.Asn167Tyr, p.Asp229Gly, p.Ala342Pro, and p.Ile406Met. Taken together, our results of functional characterization in combination with in silico prediction suggest that while p.Asn167Tyr, p.Thr200Asn, and p.Ile406Met PAH variants have a mild impact on the protein, p.Asp229Gly, p.Gly239Ala, p.Phe263Ser, and p.Ala342Pro severely affect protein structure and function.
ER  -

PECIMONOVA, Martina, Daniela KLUCKOVA, František CSICSAY, Kamila RÉBLOVÁ, Jan KRAHULEC, Dagmar PROCHÁZKOVÁ, Ludovit ŠKULTETY, Ludevít KADASI and Andrea ŠOLTYSOVÁ. Structural and Functional Impact of Seven Missense Variants of Phenylalanine Hydroxylase. \textit{Genes}. Basilej: MDPI, 2019, vol.~10, No~6, p.~1-13. ISSN~2073-4425. Available from: https://dx.doi.org/10.3390/genes10060459.

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