title: Molecular cloning and expression of pulmonary lipid phosphate phosphohydrolases.
abstract: Pulmonary lipid phosphate phosphohydrolase (LPP) was shown previously to hydrolyze phosphatidic acid and lysophosphatidic acid in purified rat lung plasma membranes. To better investigate the nature of pulmonary LPP isoforms and their role in the lung, LPPs were cloned by RT-PCR from both adult rat lung and type II cell RNA. The RT-PCR generated LPP1 (849 bp), up to three LPP1 variants, and LPP3 (936 bp) cDNAs. The three LPP1 variants include LPP1a (852 bp) and two novel isoforms, LPP1b (697 bp) and LPP1c (1004 bp). The pulmonary LPP1 and LPP3 isoforms are essentially identical to the previously cloned rat liver and intestinal LPPs, respectively, and the LPP1a isoform has 80% sequence identity to the human homolog. The LPP2 isoform was not detected in lung by RT-PCR. Northern analyses revealed that the mRNAs for LPP1 and LPP3 increase in fetal rat lung in late gestation to day 1 after birth. These mRNAs decrease somewhat during the neonatal period but increase slightly during postnatal development. Expression of LPP1, LPP1a, and LPP3 cDNAs in HEK 293 cells established that they encode functional LPP. In contrast, the novel isoforms LPP1b and LPP1c contain frameshifts that would result in premature termination, producing putative catalytically inactive polypeptides of 30 and 76 amino acids, respectively. Further investigation of the LPP1b isoform revealed that it was present across a variety of tissues, although at lower levels than LPP1/1a. Transient mammalian expression of LPP1b failed to increase phosphatidate phosphohydrolase activity in HEK 293 cells.
authors.affiliation: Medical Research Council Group in Fetal and Neonatal Health and Development, Department of Biochemistry, University of Western Ontario, London, Ontario, Canada N6A 5A5.
authors.forename: M
authors.initials: M
authors.lastname: Nanjundan
authors.forename: F
authors.initials: F
authors.lastname: Possmayer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D007668:Kidney / Q000166:cytology; D008168:Lung / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D010711:Phosphatidate Phosphatase / Q000235:genetics*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics*; D011663:Pulmonary Surfactants / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats
chemical_list: D007527:Isoenzymes; D011663:Pulmonary Surfactants; D012333:RNA, Messenger; C112948:lipid phosphate phosphatase; D010744:Phosphoric Monoester Hydrolases; D010711:Phosphatidate Phosphatase