Si. Goodman et al., CLONING OF GLUTARYL-COA DEHYDROGENASE CDNA, AND EXPRESSION OF WILD-TYPE AND MUTANT ENZYMES IN ESCHERICHIA-COLI, Human molecular genetics, 4(9), 1995, pp. 1493-1498
We have cloned, sequenced, and expressed cDNAs encoding wild type huma
n glutaryl-CoA dehydrogenase subunit, and have expressed a mutant enzy
me found in a patient with glutaric acidemia type I. The mutant protei
n is expressed at the same level as the wild type in Escherichia coil,
but has less than 1% of the activity of wild-type dehydrogenase. We a
lso present evidence that the glutaryl-CoA dehydrogenase transcript is
alternatively spliced in human fibroblasts and liver; the alternative
ly spliced mRNA, when expressed in E.coli, encodes a stable but inacti
ve protein. Purified expressed human glutaryl-CoA dehydrogenase has ki
netic constants similar to those of the previously purified porcine de
hydrogenase. The primary translation product from in vitro transcribed
glutaryl-CoA dehydrogenase mRNA is translocated into mitochondria and
processed in the same manner as most other nuclear-encoded mitochondr
ial proteins, Human glutaryl-CoA dehydrogenase shows 53% sequence simi
larity to porcine medium chain acyl-CoA dehydrogenase, and these simil
arities were utilized to predict structure-function relationships in g
lutaryl-CoA dehydrogenase.