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1995 Fiscal Year Final Research Report Summary

Propionic acidemia : bacterial expression system for propionyl CoA carboxylase.

Research Project

Project/Area Number 06670762
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Pediatrics
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

OHURA Toshihiro  TOHOKU UNIVERSITY,SCHOOL OF MEDICINE,DEPARTMENT OF PEDIATRICS,ASSISTANT PROFESSOR, 医学部, 講師 (10176828)

Co-Investigator(Kenkyū-buntansha) KURE Shigeo  TOHOKU UNIVERSITY,SCHOOL OF MEDICINE,DEPARTMENT OF BIOCHEMICAL GENETICS,CLINICAL, 医学部, 助手 (70211191)
Project Period (FY) 1994 – 1995
KeywordsPropionic acdemia / Propiony1 CoA Carboxylase / Chaperonine
Research Abstract

We developed a bacterial expression system for the human alpha and beta cDNAs of propionyl-CoA carboxylase (PCC). These cDNAs were co-expressed in E.coli on one plasmid vector with each cDNA having its own IPTG-inducible promoter. Only negligible amounts of active PCC were measured. Co-expression of this plasmid with a second plasmid vector overexpressing the E.coli chaperonin proteins, groES and groEL,resulted in a several hundred-fold increase in PCC Specific activity, to a level comparable to that found in crude human liver extracts. Deficiency of either alpha PCC or beta PCC results in propionic acidemia, an autosomal recessive disorder. We used this expression system to characterize one missense mutation previously described in five Japanese alleles, namely C1283T (Thr428Ile) in betaPCC.This mutant, when expressed in E.coli under the same conditions as that of wild type PCC,had null activity. This bacterial expression system can be useful for analysis of either alphaPCC or betaPCC mutations. Our findings indicated that the groES and groEL chaperonin proteins were essential for folding and assembly of the human PCC heteromeric subunits.

  • Research Products

    (5 results)

All Other

All Publications (5 results)

  • [Publications] 大浦 敏博: "プロピオン酸血症" Molecular Medicine臨時増刊、遺伝子病マニュアル(上). 32. 76-77 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohura T et all: "A novel splicing mutation in propionic acidemia associated with a tetranucleotide direct repeat in the PCC B gene." Human Genetics. 95. 707-708 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kelson TL et all: "Chaperonin mediated folding and overexpression of recombinant human propionyl CoA carboxylase in E coli." Am J Hum Genet. 57. A244 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohura T.et. al.: "A novel splicing mutation in propionic acidemia associated with a tetranucleotide direct repeat in the PCC B gene." Human Genetics. 95-6. 707-708 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kelson T.L.et. all.: "Chaperonin mediated folding and overexpression of recombinant human propionyl CoA carboxylase in E.coli." Am J Hum Genet. 57-4. A244 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1997-03-04  

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