Abstract
MCAD is a mitochondrial flavoprotein, a tetramer of 45 kDa subunits, which catalyzes the first reaction in the β-oxidation of medium chain fatty acids. Hereditary MCAD deficiency causes recurrent episodes of non-ketotic hypoglycemic coma, accompanied by medium chain dicarboxylic aciduria. We recently reported the isolation of partial cDNAs encoding rat and human MCAD, containing 50 and 80% of the coding region, respectively. The gene for human MCAD was assigned to chromosome 1, p31 (PNAS 83:6543, 1986). We report here the isolation and nucleotide sequence of a cDNA clone that contains the entire coding region of rat MCAD. mRNA encoding MCAD was purified from rat liver by polysome immunopurification using monospecific antibody, and a cDNA library enriched from MCAD was constructed. The library was screened by colony hybridization using a previously isolated rat MCAD cDNA clone. Numerous MCAD clones were isolated. In vitro expression of the longest clone (1860 bp) yielded a polypeptide that is indistinguishable in size from rat precursor MCAD containing the leader sequence. This peptide was immunoprecipitable with anti-MCAD antibody. The identity of the clone was further confirmed by comparison of the entire nucleotide sequence with amino acid sequences of the amino terminal and six internal peptides of rat MCAD. The leader peptide contains 25 amino acids, and is rich in basic amino acids (4 Arg, 1 Lys).
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Matsubara, Y., Kraus, J., Ozasa, H. et al. MOLECULAR CLONING AND NUCLEOTIDE SEQUENCE OF cDNA ENCODING RAT MEDIUM CHAIN ACYL-CoA DEHYDROGENASE (MCAD). Pediatr Res 21 (Suppl 4), 292 (1987). https://doi.org/10.1203/00006450-198704010-00748
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DOI: https://doi.org/10.1203/00006450-198704010-00748