OBJECTIVES: To estimate the frequency of the cystathionine beta-synthase deficiency caused by c.1105C>T mutation in Central Europe compared to Norway, and to examine the pathogenicity of the corresponding p.R369C mutant enzyme. STUDY DESIGN: Mutation c.1105C>T was analyzed in 600 anonymous Czech newborn blood spots. Catalytic activity and quaternary structure of the p.R369C mutant was evaluated after expression in 2 cellular systems. RESULTS: Population frequency of the c.1105C>T mutation was 0.005, predicting the birth prevalence of homocystinuria of 1:40000, which increased to 1:15500 in a model including 10 additional mutations. In Escherichia coli the p.R369C mutant misfolded, and its activity was severely reduced, and expression in Chinese hamster ovary cells enabled proper folding with activity decreased to 63% of the wild-type enzyme. This decreased activity was not due to impaired K(m) for both substrates but resulted from V(max) lowered to 55% of the normal cystathionine beta-synthase enzyme. CONCLUSIONS: The c.1105C>T (p.R369C) allele is common also in the Czech population. Although the p.R369C mutation impairs folding and decreases velocity of the enzymatic reaction, our data are congruent with rather mild clinical phenotype in homozygotes or compound heterozygotes carrying this mutation.

Institute of Inherited Metabolic Disorders Charles University Prague 1st Faculty of Medicine Prague Czech Republic

Zobrazit více v PubMed

Mudd S.H., Levy H.L., Kraus J.P. Disorders of transsulfuration. In: Scriver C.R., Beaudet A.L., Sly W.S., Valle D., Childs B., Vogelstein B., editors. The Metabolic and Molecular Bases of Inherited Disease. 8 ed. McGraw-Hill; New York: 2001. pp. 2007–2056.

Kraus J.P., Kozich V. Cystathionine-beta-synthase and its deficiency. In: Carmel R., Jakobsen D.W., editors. Homocysteine in Health and Disease. Cambridge University Press; Cambridge: 2001. pp. 223–243.

Gaustadnes M., Rudiger N., Rasmussen K., Ingerslev J. Familial thrombophilia associated with homozygosity for the cystathionine beta-synthase 833T—>C mutation. Arterioscler Thromb Vasc Biol. 2000;20:1392–1395. PubMed

Gaustadnes M., Wilcken B., Oliveriusova J., McGill J., Fletcher J., Kraus J.P. The molecular basis of cystathionine beta-synthase deficiency in Australian patients: genotype-phenotype correlations and response to treatment. Hum Mutat. 2002;20:117–126. PubMed

Linnebank M., Junker R., Nabavi D.G., Linnebank A., Koch H.G. Isolated thrombosis due to the cystathionine beta-synthase mutation c.833T>C (1278T) J Inherit Metab Dis. 2003;26:509–511. PubMed

Maclean K.N., Gaustadnes M., Oliveriusova J., Janosik M., Kraus E., Kozich V. High homocysteine and thrombosis without connective tissue disorders are associated with a novel class of cystathionine beta-synthase (CBS) mutations. Hum Mutat. 2002;19:641–655. PubMed

Wilcken B., Wiley V., Hammond J., Carpenter K. Screening newborns for inborn errors of metabolism by tandem mass spectrometry. N Engl J Med. 2003;348:2304–2312. PubMed

Naughten E.R., Yap S., Mayne P.D. Newborn screening for homocystinuria: Irish and world experience. Eur J Pediatr. 1998;157(Suppl 2):S84–S87. PubMed

Gaustadnes M., Ingerslev J., Rutiger N. Prevalence of congenital homocystinuria in Denmark. N Engl J Med. 1999;340:1513. PubMed

Sokolova J., Janosikova B., Terwilliger J.D., Freiberger T., Kraus J.P., Kozich V. Cystathionine beta-synthase deficiency in Central Europe: discrepancy between biochemical and molecular genetic screening for homocystinuric alleles. Hum Mutat. 2001;18:548–549. PubMed

Linnebank M., Homberger A., Junker R., Nowak-Goettl U., Harms E., Koch H.G. High prevalence of the I278T mutation of the human cystathionine beta-synthase detected by a novel screening application. Thromb Haemost. 2001;85:986–988. PubMed

Refsum H., Fredriksen A., Meyer K., Ueland P.M., Kase B.F. Birth prevalence of homocystinuria. J Pediatr. 2004;144:830–832. PubMed

Kluijtmans L.A., Boers G.H., Kraus J.P., van den Heuvel L.P., Cruysberg J.R., Trijbels F.J. The molecular basis of cystathionine beta-synthase deficiency in Dutch patients with homocystinuria: effect of CBS genotype on biochemical and clinical phenotype and on response to treatment. Am J Hum Genet. 1999;65:59–67. PubMed PMC

Kim C.E., Gallagher P.M., Guttormsen A.B., Refsum H., Ueland P.M., Ose L. Functional modeling of vitamin responsiveness in yeast: a common pyridoxine-responsive cystathionine beta-synthase mutation in homocystinuria. Hum Mol Genet. 1997;6:2213–2221. PubMed

Kozich V., Kraus J.P. Screening for mutations by expressing patient cDNA segments in E. coli: homocystinuria due to cystathionine beta-synthase deficiency. Hum Mutat. 1992;1:113–123. PubMed

Lowry O.H., Rosenbrough N.J., Farr A.L., Randall R.J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–275. PubMed

Kraus J.P. Cystathionine beta-synthase (human) Methods Enzymol. 1987;143:388–394. PubMed

Meier M., Oliveriusova J., Kraus J.P., Burkhard P. Structural insights into mutations of cystathionine beta-synthase. Biochim Biophys Acta. 2003;1647:206–213. PubMed

Yamanishi M., Kabil O., Sen S., Banerjee R. Structural insights into pathogenic mutations in heme-dependent cystathionine-beta-synthase. J Inorg Biochem. 2006;100:1988–1995. PubMed

Brown C.R., Hong-Brown L.Q., Welch W.J. Correcting temperature-sensitive protein folding defects. J Clin Invest. 1997;99:1432–1444. PubMed PMC

Waters P.J. How PAH gene mutations cause hyper-phenylalaninemia and why mechanism matters: insights from in vitro expression. Hum Mutat. 2003;21:357–369. PubMed

Mandey S.H., Schneiders M.S., Koster J., Waterham H.R. Mutational spectrum and genotype-phenotype correlations in mevalonate kinase deficiency. Hum Mutat. 2006;27:796–802. PubMed

Peterschmitt M.J., Simmons J.R., Levy H.L. Reduction of false negative results in screening of newborns for homocystinuria. N Engl J Med. 1999;341:1572–1576. PubMed

Refsum H., Grindflek A.W., Ueland P.M., Fredriksen A., Meyer K., Ulvik A. Screening for serum total homocysteine in newborn children. Clin Chem. 2004;50:1769–1784. PubMed

Bowron A., Barton A., Scott J., Stansbie D. Blood spot homocysteine: a feasibility and stability study. Clin Chem. 2005;51:257–258. PubMed

Linnebank M., Homberger A., Kraus J.P., Harms E., Kozich V., Koch H.G. Haplotyping of wild-type and I278T alleles of the human cystathionine beta-synthase gene based on a cluster of novel SNPs in IVS12. Hum Mutat. 2001;17:350–351. PubMed

A proactive genotype-to-patient-phenotype map for cystathionine beta-synthase

Guidelines for the diagnosis and management of cystathionine beta-synthase deficiency

Newborn screening for homocystinurias and methylation disorders: systematic review and proposed guidelines

Conformational properties of nine purified cystathionine β-synthase mutants

Determination of cystathionine beta-synthase activity in human plasma by LC-MS/MS: potential use in diagnosis of CBS deficiency

Vascular presentation of cystathionine beta-synthase deficiency in adulthood

Restoring assembly and activity of cystathionine β-synthase mutants by ligands and chemical chaperones

Cystathionine beta-synthase mutations: effect of mutation topology on folding and activity