In silico and in vivo models for Qatari-specific classical homocystinuria as basis for development of novel therapies

Hesham M. Ismail, Navaneethakrishnan Krishnamoorthy, Nader Al-Dewik, Hatem Zayed, Nura A. Mohamed, Valeria Di Giacomo, Sapna Gupta, Johannes Häberle, Beat Thöny, Henk J. Blom, Waren D. Kruger, Tawfeg Ben-Omran, Gheyath K. Nasrallah

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1 Citation (Scopus)


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. Wild-type 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 reduced 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.

Original languageEnglish
JournalHuman Mutation
Publication statusAccepted/In press - 1 Jan 2018



  • CBS
  • chemical chaperones
  • Homocystinuria
  • in silico
  • in vivo models
  • p.R336C mutation
  • Qatar

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Ismail, H. M., Krishnamoorthy, N., Al-Dewik, N., Zayed, H., Mohamed, N. A., Giacomo, V. D., Gupta, S., Häberle, J., Thöny, B., Blom, H. J., Kruger, W. D., Ben-Omran, T., & Nasrallah, G. K. (Accepted/In press). In silico and in vivo models for Qatari-specific classical homocystinuria as basis for development of novel therapies. Human Mutation.