During the past 20 years, cystathionine beta-synthase (CBS) deficiency has been detected in the former Czechoslovakia with a calculated frequency of 1:349,000. The clinical manifestation was typical of homocystinuria, and about half of the 21 patients were not responsive to pyridoxine. Twelve distinct mutations were detected in 30 independent homocystinuric alleles. One half of the alleles carried either the c.833 T-->C or the IVS11-2A-->C mutation; the remaining alleles contained private mutations. The abundance of five mutant mRNAs with premature stop codons was analyzed by PCR-RFLP. Two mRNAs, c.828_931ins104 (IVS7+1G-->A) and c.1226 G-->A, were severely reduced in the cytoplasm as a result of nonsense-mediated decay. In contrast, the other three mRNAs-c.19_20insC, c.28_29delG, and c.210_235del26 (IVS1-1G-->C)-were stable. Native western blot analysis of 14 mutant fibroblast lines showed a paucity of CBS antigen, which was detectable only in aggregates. Five mutations-A114V (c.341C-->T), A155T (c.463G-->A), E176K (c.526G-->A), I278T (c.833T-->C), and W409_G453del (IVS11-2A-->C)-were expressed in Escherichia coli. All five mutant proteins formed substantially more aggregates than did the wild-type CBS, and no aggregates contained heme. These data suggest that abnormal folding, impaired heme binding, and aggregation of mutant CBS polypeptides may be common pathogenic mechanisms in CBS deficiency.

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

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Authors' Web site, http://www.uchsc.edu/sm/cbs (for an updated list of mutations)

Genbank, http://www.ncbi.nlm.nih.gov/Genbank (for human CBS cDNA [accession number L19501] and genomic DNA [accession number AF042836])

Online Mendelian Inheritance in Man (OMIM), http://www.ncbi.nlm.nih.gov/Omim/ (for CBS deficiency [MIM 236200])

Primers and Conditions, http://www.lf1.cuni.cz/~mjano/protocols.html (for list of PCR primers and conditions for amplification of all 23 CBS exons from genomic DNA)

Bross P, Corydon TJ, Andresen BS, Jorgensen MM, Bolund L, Gregersen N (1999) Protein misfolding and degradation in genetic diseases. Hum Mutat 14:186–198 PubMed

Cooper DN, Krawczak M, Antonarakis SE (1995) The nature and mechanisms of human gene mutation. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited disease. 7th ed. McGraw-Hill, New York, pp 259–292

Dawson PA, Cox AJ, Emmerson BT, Dudman NP, Kraus JP, Gordon RB (1997) Characterisation of five missense mutations in the cystathionine beta-synthase gene from three patients with B6-nonresponsive homocystinuria. Eur J Hum Genet 5:15–21 PubMed

de Franchis R, Kozich V, McInnes RR, Kraus JP (1994) Identical genotypes in siblings with different homocystinuric phenotypes: identification of three mutations in cystathionine beta-synthase using an improved bacterial expression system. Hum Mol Genet 3:1103–1108 PubMed

Farabaugh PJ (2000) Translational frameshifting: implications for the mechanism of translational frame maintenance. Prog Nucleic Acid Res Mol Biol 64:131–170 PubMed

Frischmeyer PA, Dietz HC (1999) Nonsense-mediated mRNA decay in health and disease. Hum Mol Genet 8:1893–1900 PubMed

Horsburgh BC, Kollmus H, Hauser H, Coen DM (1996) Translational recoding induced by G-rich mRNA sequences that form unusual structures. Cell 86:949–959 PubMed PMC

Johnston JA, Ward CL, Kopito RR (1998) Aggresomes: a cellular response to misfolded proteins. J Cell Biol 143:1883–1898 PubMed PMC

Kery V, Bukovska G, Kraus JP (1994) Transsulfuration depends on heme in addition to pyridoxal 5′-phosphate: cystathionine beta-synthase is a heme protein. J Biol Chem 269:25283–288 PubMed

Kery V, Elleder D, Kraus JP (1995) Delta-aminolevulinate increases heme saturation and yield of human cystathionine beta-synthase expressed in Escherichia coli. Arch Biochem Biophys 316:24–29 PubMed

Kery V, Poneleit L, Elleder D, Straubhaar JR, Teisinger J, Kraus JP (1997) The carboxy terminus of human cystathionine β-synthase is involved in tetramer formation and activation of the enzyme by S-adenosyl-l-methionine. Faseb J 11:1613

Kery V, Poneleit L, Meyer JD, Manning MC, Kraus JP (1999) Binding of pyridoxal 5′-phosphate to the heme protein human cystathionine beta-synthase. Biochemistry 38:2716–2724 PubMed

Kirschner M (1999) Intracellular proteolysis. Trends Cell Biol 9:M42–45 PubMed

Kluijtmans LA, Boers GH, Kraus JP, van den Heuvel LP, Cruysberg JR, Trijbels FJ, Blom HJ (1999) 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 65:59–67 PubMed PMC

Koch HG, Ullrich K, Deufel T, Harms E (1994) A highly prevalent splice site mutation in the cystathionine-β-synthase gene is associated with poor vitamin B6 response in homocystinuria. Pediatr Res 36:21A

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

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

Kraus JP, Janosik M, Kozich V, Mandell R, Shih V, Sperandeo MP, Sebastio G, de Franchis R, Andria G, Kluijtmans LA, Blom H, Boers GH, Gordon RB, Kamoun P, Tsai MY, Kruger WD, Koch HG, Ohura T, Gaustadnes M (1999) Cystathionine beta-synthase mutations in homocystinuria. Hum Mutat 13:362–375 PubMed

Kraus JP, Le K, Swaroop M, Ohura T, Tahara T, Rosenberg LE, Roper MD, Kozich V (1993) Human cystathionine beta-synthase cDNA: sequence, alternative splicing and expression in cultured cells. Hum Mol Genet 2:1633–1638 PubMed

Mudd SH, Levy HL, Kraus JP (2001) Disorders of transsulfuration. In: Scriver CR, Beaudet AL, Sly WS, Valle D, Childs B, Vogelstein B (eds) The metabolic and molecular bases of inherited disease. 8th ed. McGraw-Hill, New York, pp 2007–2056

Mudd SH, Skovby F, Levy HL, Pettigrew KD, Wilcken B, Pyeritz RE, Andria G, Boers GH, Bromberg IL, Cerone R, Fowler B, Grobe H, Schmidt H, Schweitzer L (1985) The natural history of homocystinuria due to cystathionine beta-synthase deficiency. Am J Hum Genet 37:1–31 PubMed PMC

Nagy E, Maquat LE (1998) A rule for termination-codon position within intron-containing genes: when nonsense affects RNA abundance. Trends Biochem Sci 23:198–199 PubMed

Shan X, Kruger WD (1998) Correction of disease-causing CBS mutations in yeast. Nat Genet 19:91–93 PubMed

Skovby F, Kraus JP, Rosenberg LE (1984) Homocystinuria: biogenesis of cystathionine beta-synthase subunits in cultured fibroblasts and in an in vitro translation system programmed with fibroblast messenger RNA. Am J Hum Genet 36:452–459 PubMed PMC

Vargas C, McEwan AG, Downie JA (1993) Detection of c-type cytochromes using enhanced chemiluminescence. Anal Biochem 209:323–326 PubMed

Zhang J, Maquat LE (1997) Evidence that translation reinitiation abrogates nonsense-mediated mRNA decay in mammalian cells. Embo J 16:826–833 PubMed PMC

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

Chaperone therapy for homocystinuria: the rescue of CBS mutations by heme arginate

Conformational properties of nine purified cystathionine β-synthase mutants

Cystathionine beta-synthase mutants exhibit changes in protein unfolding: conformational analysis of misfolded variants in crude cell extracts

Novel structural arrangement of nematode cystathionine β-synthases: characterization of Caenorhabditis elegans CBS-1

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

Diversity of cystathionine beta-synthase haplotypes bearing the most common homocystinuria mutation c.833T>C: a possible role for gene conversion

Homocystinuria due to cystathionine beta-synthase deficiency: novel biochemical findings and treatment efficacy

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GENBANK
AF042836, L19501

OMIM
236200