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. 1985 Mar 25;13(6):1891–1903. doi: 10.1093/nar/13.6.1891

Molecular cloning and nucleotide sequence of rat lingual lipase cDNA.

A J Docherty, M W Bodmer, S Angal, R Verger, C Riviere, P A Lowe, A Lyons, J S Emtage, T J Harris
PMCID: PMC341123  PMID: 3839077

Abstract

Purified rat lingual lipase (EC3113), a glycoprotein of approximate molecular weight 52,000, was used to generate polyclonal antibodies which were able to recognise the denatured and deglycosylated enzyme. These immunoglobulins were used to screen a cDNA library prepared from mRNA isolated from the serous glands of rat tongue cloned in E. coli expression vectors. An almost full length cDNA clone was isolated and the nucleotide and predicted amino acid sequence obtained. Comparison with the N-terminal amino acid sequence of the purified enzyme confirmed the identity of the cDNA and indicated that there was a hydrophobic signal sequence of 18 residues. The amino acid sequence of mature rat lingual lipase consists of 377 residues and shares little homology with porcine pancreatic lipase apart from a short region containing a serine residue at an analogous position to the ser 152 of the porcine enzyme.

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Selected References

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  1. Abrams C. K., Hamosh M., Hubbard V. S., Dutta S. K., Hamosh P. Lingual lipase in cystic fibrosis. Quantitation of enzyme activity in the upper small intestine of patients with exocrine pancreatic insufficiency. J Clin Invest. 1984 Feb;73(2):374–382. doi: 10.1172/JCI111222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benyajati C., Spoerel N., Haymerle H., Ashburner M. The messenger RNA for alcohol dehydrogenase in Drosophila melanogaster differs in its 5' end in different developmental stages. Cell. 1983 May;33(1):125–133. doi: 10.1016/0092-8674(83)90341-0. [DOI] [PubMed] [Google Scholar]
  4. Brown A. J., Leibold E. A., Munro H. N. Isolation of cDNA clones for the light subunit of rat liver ferritin: evidence that the light subunit is encoded by a multigene family. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1265–1269. doi: 10.1073/pnas.80.5.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  6. Carey M. C., Small D. M., Bliss C. M. Lipid digestion and absorption. Annu Rev Physiol. 1983;45:651–677. doi: 10.1146/annurev.ph.45.030183.003251. [DOI] [PubMed] [Google Scholar]
  7. Chapus C., Sémériva M. Mechanism of pancreatic lipase action. 2. Catalytic properties of modified lipases. Biochemistry. 1976 Nov 16;15(23):4988–4991. doi: 10.1021/bi00668a007. [DOI] [PubMed] [Google Scholar]
  8. Cheng Y. S. Increased cell buoyant densities of protein overproducing Escherichia coli cells. Biochem Biophys Res Commun. 1983 Feb 28;111(1):104–111. doi: 10.1016/s0006-291x(83)80123-5. [DOI] [PubMed] [Google Scholar]
  9. Field R. B., Scow R. O. Purification and characterization of rat lingual lipase. J Biol Chem. 1983 Dec 10;258(23):14563–14569. [PubMed] [Google Scholar]
  10. Guidoni A., Benkouka F., De Caro J., Rovery M. Characterization of the serine reacting with diethyl p-nitrophenyl phosphate in porcine pancreatic lipase. Biochim Biophys Acta. 1981 Jul 24;660(1):148–150. doi: 10.1016/0005-2744(81)90120-0. [DOI] [PubMed] [Google Scholar]
  11. Hamosh M., Burns W. A. Lipolytic activity of human lingual glands (Ebner). Lab Invest. 1977 Dec;37(6):603–608. [PubMed] [Google Scholar]
  12. Hamosh M., Scow R. O. Lingual lipase and its role in the digestion of dietary lipid. J Clin Invest. 1973 Jan;52(1):88–95. doi: 10.1172/JCI107177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  14. Harris T. J., Brown F., Sangar D. V. Differential precipitation of foot and mouth disease virus proteins made in vivo and in vitro by hyperimmune and virus particle guinea pig antisera. Virology. 1981 Jul 15;112(1):91–98. doi: 10.1016/0042-6822(81)90615-2. [DOI] [PubMed] [Google Scholar]
  15. Helfman D. M., Feramisco J. R., Fiddes J. C., Thomas G. P., Hughes S. H. Identification of clones that encode chicken tropomyosin by direct immunological screening of a cDNA expression library. Proc Natl Acad Sci U S A. 1983 Jan;80(1):31–35. doi: 10.1073/pnas.80.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kemp D. J., Coppel R. L., Cowman A. F., Saint R. B., Brown G. V., Anders R. F. Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: detection with antibodies from immune humans. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3787–3791. doi: 10.1073/pnas.80.12.3787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Mellor J., Dobson M. J., Roberts N. A., Tuite M. F., Emtage J. S., White S., Lowe P. A., Patel T., Kingsman A. J., Kingsman S. M. Efficient synthesis of enzymatically active calf chymosin in Saccharomyces cerevisiae. Gene. 1983 Sep;24(1):1–14. doi: 10.1016/0378-1119(83)90126-9. [DOI] [PubMed] [Google Scholar]
  20. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  21. Müller-Hill B., Crapo L., Gilbert W. Mutants that make more lac repressor. Proc Natl Acad Sci U S A. 1968 Apr;59(4):1259–1264. doi: 10.1073/pnas.59.4.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Patel T. P., Millican T. A., Bose C. C., Titmas R. C., Mock G. A., Eaton M. A. Improvements to solid phase phosphotriester synthesis of deoxyoligonucleotides. Nucleic Acids Res. 1982 Sep 25;10(18):5605–5620. doi: 10.1093/nar/10.18.5605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  24. Retzel E. F., Collett M. S., Faras A. J. Enzymatic synthesis of deoxyribonucleic acid by the avian retrovirus reverse transcriptase in vitro: optimum conditions required for transcription of large ribonucleic acid templates. Biochemistry. 1980 Feb 5;19(3):513–518. doi: 10.1021/bi00544a019. [DOI] [PubMed] [Google Scholar]
  25. Ricciardi R. P., Miller J. S., Roberts B. E. Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4927–4931. doi: 10.1073/pnas.76.10.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rose J. K., Gallione C. J. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus G and M proteins determined from cDNA clones containing the complete coding regions. J Virol. 1981 Aug;39(2):519–528. doi: 10.1128/jvi.39.2.519-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rouard M., Sari H., Nurit S., Entressangles B., Desnuelle P. Inhibition of pancreatic lipase by mixed micelles of diethyl p-nitrophenyl phosphate and bile salts. Biochim Biophys Acta. 1978 Aug 25;530(2):227–235. doi: 10.1016/0005-2760(78)90008-5. [DOI] [PubMed] [Google Scholar]
  28. Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Suggs S. V., Wallace R. B., Hirose T., Kawashima E. H., Itakura K. Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6613–6617. doi: 10.1073/pnas.78.11.6613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tiruppathi C., Balasubramanian K. A. Purification and properties of an acid lipase from human gastric juice. Biochim Biophys Acta. 1982 Sep 14;712(3):692–697. doi: 10.1016/0005-2760(82)90299-5. [DOI] [PubMed] [Google Scholar]
  33. Verger R., de Haas G. H., Sarda L., Desnuelle Purification from porcine pancreas of two molecular species with lipase activity. Biochim Biophys Acta. 1969;188(2):272–282. doi: 10.1016/0005-2795(69)90075-0. [DOI] [PubMed] [Google Scholar]
  34. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  35. Watson M. E. Compilation of published signal sequences. Nucleic Acids Res. 1984 Jul 11;12(13):5145–5164. doi: 10.1093/nar/12.13.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Williams D. C., Van Frank R. M., Muth W. L., Burnett J. P. Cytoplasmic inclusion bodies in Escherichia coli producing biosynthetic human insulin proteins. Science. 1982 Feb 5;215(4533):687–689. doi: 10.1126/science.7036343. [DOI] [PubMed] [Google Scholar]
  37. de Caro J., Boudouard M., Bonicel J., Guidoni A., Desnuelle P., Rovery M. Porcine pancreatic lipase. Completion of the primary structure. Biochim Biophys Acta. 1981 Dec 29;671(2):129–138. doi: 10.1016/0005-2795(81)90126-4. [DOI] [PubMed] [Google Scholar]

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