Laboratory of Cellular and Molecular Pharmacology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709.
Preparations of mRNA isolated from rabbit lung and liver were used in the construction of libraries that were screened for cDNAs encoding the pulmonary or hepatic isozyme of the flavin-containing monooxygenase. The hepatic library was screened with cDNA encoding the flavin-containing monooxygenase expressed in pig liver, and a clone containing a 2.0-kilobase insert was detected and isolated. This cDNA insert encoded a protein of 535 amino acids with a primary structure 87% identical to that of the pig flavin-containing monooxygenase. The pulmonary library was screened with polyclonal antibodies to the flavin-containing monooxygenase expressed in rabbit lung, and a clone containing a 2.6-kilobase insert was detected and isolated. Although the protein encoded by this insert also contained 535 amino acids, its primary sequence was only 56% identical to that of the liver enzyme. The sequences of several peptides obtained by digestion of the purified rabbit pulmonary flavin-containing monooxygenase with trypsin matched exactly with sequences derived from the cDNA structure. Tissue-specific distribution of mRNA for the hepatic and pulmonary isozymes of the flavin-containing monooxygenase was consistent with the distribution of protein, an indication that expression of flavin-containing monooxygenase is controlled at the level of transcription. Analysis of genomic DNA indicates that both the hepatic and pulmonary enzymes may be products of single genes.