Back to Search
Start Over
In silico and in vivo models for Qatari-specific classical homocystinuria as basis for development of novel therapies
- Publication Year :
- 2018
-
Abstract
- Homocystinuria is a rare inborn error of methionine metabolism caused by cystathionine β-synthase (CBS) deficiency. The prevalence of homocystinuria in Qatar is 1:1,800 births, mainly due to a founder Qatari missense mutation, c.1006C>T; p.R336C (p.Arg336Cys). We characterized the structure-function relationship of the p.R336C mutant protein, and investigated the effect of different chemical chaperones to restore p.R336C-CBS activity using three models: In silico, ΔCBS yeast, and CRISPR/Cas9 p.R336C knock-in HEK293T and HepG2 cell lines. Protein modeling suggested that the p.R336C induces severe conformational and structural changes, perhaps influencing CBS activity. Wildtype CBS, but not the p.R336C mutant, was able to restore the yeast growth in ΔCBS deficient yeast in a complementation assay. The p.R336C knock-in HEK293T and HepG2 cells decreased the level of CBS expression and reduce its structural stability; however, treatment of the p.R336C knock-in HEK293T cells with betaine, a chemical chaperone, restored the stability and tetrameric conformation of CBS, but not its activity. Collectively, these results indicate that the p.R336C mutation has a deleterious effect on CBS structure, stability, and activity, and using the chemical chaperones approach for treatment could be ineffective in restoring p.R336C CBS activity. This article is protected by copyright. All rights reserved. QNRF
- Subjects :
- Protein Folding
congenital, hereditary, and neonatal diseases and abnormalities
2716 Genetics (clinical)
In silico
Mutant
Mutation, Missense
p.R336C mutation
Cystathionine beta-Synthase
Homocystinuria
610 Medicine & health
medicine.disease_cause
Gene Expression Regulation, Enzymologic
Article
CBS
Structure-Activity Relationship
03 medical and health sciences
Methionine
1311 Genetics
Protein-fragment complementation assay
Enzyme Stability
Genetics
medicine
Humans
Missense mutation
Computer Simulation
Qatar
Genetics (clinical)
chemical chaperones
030304 developmental biology
0303 health sciences
Mutation
biology
030305 genetics & heredity
nutritional and metabolic diseases
Hep G2 Cells
medicine.disease
Cystathionine beta synthase
Protein Structure, Tertiary
HEK293 Cells
Biochemistry
in silico
10036 Medical Clinic
biology.protein
in vivo models
Mutant Proteins
Chemical chaperone
Molecular Chaperones
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....9553df58aa8ecfb89c3db22f01c5d601