NUCLEOTIDE-SEQUENCE ENCODING A NOVEL MEMBER OF THE HYDRATASE DEHYDROGENASE FAMILY

The nucleotide sequence encoding a novel member of the family of fatty acid oxidation enzymes has been determined. Clones were generated fro m porcine gastric corpus cDNA by application of the polymerase chain r eaction (PCR) using oligodeoxyribonucleotides based on the amino acid sequence of a 78 kDa gastrin-binding protein isolated from porcine gas tric mucosal membranes (Baldwin et al., J. Biol. Chem. 261 (1986) 1225 2-12257). Clones encoding the 3' and 5' ends of the cDNA were then iso lated by conventional screening of a porcine liver cDNA library, and b y application of anchored PCR, respectively. The composite cDNA of 274 4 nucleotides encoded a protein of 763 amino acids, the sequence of wh ich was related to a rat peroxisomal trifunctional enzyme, DELTA3, DEL TA2-enoyl-CoA isomerase/enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrog enase, and to other members of the same enzyme family. Northern blots indicated that the 3-kb mRNA encoding the novel protein was abundant i n gastric corpus and liver, with lower amounts also present in gastric antrum and forestomach.