Impaired heme binding and aggregation of mutant cystathionine beta-synthase subunits in homocystinuria

Am J Hum Genet. 2001 Jun;68(6):1506-13. doi: 10.1086/320597. Epub 2001 May 15.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adolescent
  • Adult
  • Alleles
  • Blotting, Western
  • Child
  • Codon, Nonsense / genetics
  • Codon, Terminator / genetics
  • Cystathionine beta-Synthase / chemistry
  • Cystathionine beta-Synthase / deficiency
  • Cystathionine beta-Synthase / genetics*
  • Cystathionine beta-Synthase / metabolism*
  • Female
  • Fibroblasts
  • Genotype
  • Heme / metabolism*
  • Homocystinuria / enzymology*
  • Homocystinuria / genetics*
  • Homocystinuria / metabolism
  • Humans
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Mutation / genetics*
  • Mutation, Missense / genetics
  • Polymorphism, Restriction Fragment Length
  • Protein Binding
  • Protein Subunits
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism


  • Codon, Nonsense
  • Codon, Terminator
  • Protein Subunits
  • RNA, Messenger
  • Heme
  • Cystathionine beta-Synthase

Associated data

  • GENBANK/AF042836
  • GENBANK/L19501
  • OMIM/236200