1995 Fiscal Year Final Research Report Summary
Molecular basis of neonatal-onset multiple carboxylase deficiency
| Project/Area Number |
05454282
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
NARISAWA Kuniaki TOHOKU UNIVERSITY,BIOCHEMICAL GENETICS,PROFESSOR, 医学部, 教授 (90004647)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Yoichi TOHOKU UNIVERSITY,BIOCHEMICAL GENETICS,ASSISTANT, 医学部, 助手 (80216457)
KURE Shigeo TOHOKU UNIVERSITY,BIOCHEMICAL GENETICS,ASSISTANT, 医学部, 助手 (10205221)
MATSUBARA Yoichi TOHOKU UNIVERSITY,BIOCHEMICAL GENETICS,ASSOCIATE PROFESSOR, 医学部, 助教授 (00209602)
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| Project Period (FY) |
1993 – 1995
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| Keywords | multiple carboxylase deficiency / biotine / holocarboxylase synthetase / HAS / holocarboxylase synthetase deficiency / HCScDNA / mutation / genetic diagnosis |
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| Research Abstract |
Neonatal-onset multiple carboxylase deficiency is known to be caused by deficiency of holocarboxylase synthetase (HCS). HCS plays an essential role in biotin utilization in cells. We purified HCS from bovine liver cytosol and found that it consists of a single subunit with a molecular mass of 64,000 Da. Identification of this polypeptide from bovine liver enabled us to determine the partial amino acid sequence of eukaryotic HCS.We have cloned the human HCS cDNA,which maps to chromosome 21q22.1. Two mutations in the HCS genes of Japanese patients with HCS deficiency have been identified : a transition from T to C which causes an amino acid substitution of proline for leucine at position 237 (L237P) and A single guanine base deletion (DELTAG1067) followed by premature termination. Transient expression analysis in cultured fibroblasts from a patient after site-directed mutagenesis demonstrated that the L237P mutant construct produce markedly decreased HCS activity, despite the presence of normal amounts ofimmunoreactive protein by Western blotting. Hybridization analysis using allele-specific oligonucleotide probes demonstrated that the prevalance of the mutations-L237P and DELTAG1067-was 50% and 30%, respectively, among Japanese patients with HCS deficiency.
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Research Products
(12 results)
