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. 1984 Jan;81(2):313–317. doi: 10.1073/pnas.81.2.313

Cloning of a cDNA encoding rat intestinal fatty acid binding protein.

D H Alpers, A W Strauss, R K Ockner, N M Bass, J I Gordon
PMCID: PMC344666  PMID: 6582489

Abstract

Intestinal fatty acid binding protein mRNA is one of the most abundant mRNA species in the rat small intestinal epithelium. RNA transfer blot analyses disclosed that the mRNA encoding intestinal fatty acid binding protein is approximately equal to 900 nucleotides long and not represented in liver RNA. We have identified 564 nucleotides of this mRNA, including 12 nucleotides of the 5' nontranslated region, the coding portion, and 155 nucleotides of the 3' nontranslated domain. The primary translation product encoded by this mRNA contains 132 amino acids and has a Mr of 15,062. The derived protein sequence was verified by automated sequential Edman degradation of the intact polypeptide isolated from a wheat germ cell-free system. The in vitro product is NH2-terminally acetylated, a finding that is consistent with its ultimate cytoplasmic destination. Comparison of the amino acid sequence of this protein with liver fatty acid binding protein, a polypeptide specified by the most abundant small intestinal epithelial mRNA, revealed significant homology and similarity in the predicted secondary structures of their NH2-terminal domains.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  3. Billheimer J. T., Gaylor J. L. Cytosolic modulators of activities of microsomal enzyme of cholesterol biosynthesis. Role of a cytosolic protein with properties similar to Z-protein (fatty acid-binding protein). J Biol Chem. 1980 Sep 10;255(17):8128–8135. [PubMed] [Google Scholar]
  4. Chan S. J., Ackerman E. J., Quinn P. S., Sigler P. B., Steiner D. F. Use of formylated yeast initiator Met tRNA to define the NH2-terminal residues of rat preproinsulin and pregrowth hormone. J Biol Chem. 1981 Apr 10;256(7):3271–3275. [PubMed] [Google Scholar]
  5. Chen-Kiang S., Wolgemuth D. J., Hsu M. T., Darnell J. E., Jr Transcription and accurate polyadenylation in vitro of RNA from the major late adenovirus 2 transcription unit. Cell. 1982 Mar;28(3):575–584. doi: 10.1016/0092-8674(82)90212-4. [DOI] [PubMed] [Google Scholar]
  6. Chou P. Y., Fasman G. D. Conformational parameters for amino acids in helical, beta-sheet, and random coil regions calculated from proteins. Biochemistry. 1974 Jan 15;13(2):211–222. doi: 10.1021/bi00699a001. [DOI] [PubMed] [Google Scholar]
  7. Deeley R. G., Gordon J. I., Burns A. T., Mullinix K. P., Binastein M., Goldberg R. F. Primary activation of the vitellogenin gene in the rooster. J Biol Chem. 1977 Nov 25;252(22):8310–8319. [PubMed] [Google Scholar]
  8. Dempsey M. E., McCoy K. E., Baker H. N., Dimitriadou-Vafiadou A., Lorsbach T., Howard J. B. Large scale purification and structural characterization of squalene and sterol carrier protein. J Biol Chem. 1981 Feb 25;256(4):1867–1873. [PubMed] [Google Scholar]
  9. Gangl A., Ockner R. K. Intestinal metabolism of plasma free fatty acids. Intracellular compartmentation and mechanisms of control. J Clin Invest. 1975 Apr;55(4):803–813. doi: 10.1172/JCI107991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gordon J. I., Alpers D. H., Ockner R. K., Strauss A. W. The nucleotide sequence of rat liver fatty acid binding protein mRNA. J Biol Chem. 1983 Mar 10;258(5):3356–3363. [PubMed] [Google Scholar]
  11. Gordon J. I., Burns A. T., Christmann J. L., Deeley R. G. Cloning of a double-stranded cDNA that codes for a portion of chicken preproalbumin. A general method for isolating a specific DNA sequence from partially purified mRNA. J Biol Chem. 1978 Dec 10;253(23):8629–8639. [PubMed] [Google Scholar]
  12. Gordon J. I., Smith D. P., Alpers D. H., Strauss A. W. Cloning of a complementary deoxyribonucleic acid encoding a portion of rat intestinal preapolipoprotein AIV messenger ribonucleic acid. Biochemistry. 1982 Oct 26;21(22):5424–5431. doi: 10.1021/bi00265a007. [DOI] [PubMed] [Google Scholar]
  13. Gordon J. I., Smith D. P., Alpers D. H., Strauss A. W. Proteolytic processing of the primary translation product of rat intestinal apolipoprotein A-IV mRNA. Comparison with preproapolipoprotein A-I processing. J Biol Chem. 1982 Jul 25;257(14):8418–8423. [PubMed] [Google Scholar]
  14. Gordon J. I., Smith D. P., Andy R., Alpers D. H., Schonfeld G., Strauss A. W. The primary translation product of rat intestinal apolipoprotein A-I mRNA is an unusual preproprotein. J Biol Chem. 1982 Jan 25;257(2):971–978. [PubMed] [Google Scholar]
  15. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HOFMANN A. F., BORGSTROEM B. THE INTRALUMINAL PHASE OF FAT DIGESTION IN MAN: THE LIPID CONTENT OF THE MICELLAR AND OIL PHASES OF INTESTINAL CONTENT OBTAINED DURING FAT DIGESTION AND ABSORPTION. J Clin Invest. 1964 Feb;43:247–257. doi: 10.1172/JCI104909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ketterer B., Tipping E., Hackney J. F., Beale D. A low-molecular-weight protein from rat liver that resembles ligandin in its binding properties. Biochem J. 1976 Jun 1;155(3):511–521. doi: 10.1042/bj1550511. [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. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  21. McCarty K. S., Jr, Vollmer R. T., McCarty K. S. Improved computer program data for the resolution and fractionation of macromolecules by isokinetic sucrose density gradient sedimentation. Anal Biochem. 1974 Sep;61(1):165–183. doi: 10.1016/0003-2697(74)90343-1. [DOI] [PubMed] [Google Scholar]
  22. Mumford R. A., Pickett C. B., Zimmerman M., Strauss A. W. Protease activities present in wheat germ and rabbit reticulocyte lysates. Biochem Biophys Res Commun. 1981 Nov 30;103(2):565–572. doi: 10.1016/0006-291x(81)90489-7. [DOI] [PubMed] [Google Scholar]
  23. O'Doherty P. J., Kuksis A. Stimulation of triacylglycerol synthesis by Z protein in rat liver and intestinal mucosa. FEBS Lett. 1975 Dec 15;60(2):256–258. doi: 10.1016/0014-5793(75)80725-3. [DOI] [PubMed] [Google Scholar]
  24. Ockner R. K., Manning J. A. Fatty acid-binding protein in small intestine. Identification, isolation, and evidence for its role in cellular fatty acid transport. J Clin Invest. 1974 Aug;54(2):326–338. doi: 10.1172/JCI107768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ockner R. K., Manning J. A., Kane J. P. Fatty acid binding protein. Isolation from rat liver, characterization, and immunochemical quantification. J Biol Chem. 1982 Jul 10;257(13):7872–7878. [PubMed] [Google Scholar]
  26. Ockner R. K., Manning J. A., Poppenhausen R. B., Ho W. K. A binding protein for fatty acids in cytosol of intestinal mucosa, liver, myocardium, and other tissues. Science. 1972 Jul 7;177(4043):56–58. doi: 10.1126/science.177.4043.56. [DOI] [PubMed] [Google Scholar]
  27. Palmiter R. D. Prevention of NH2-terminal acetylation of proteins synthesized in cell-free systems. J Biol Chem. 1977 Dec 25;252(24):8781–8783. [PubMed] [Google Scholar]
  28. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Schechter I., Wolf O., Kantor F., Schechter B., Burstein Y. Immunoglobulin precursors: structure, function, gene-protein correlation and evolution. Ann N Y Acad Sci. 1980;343:218–231. doi: 10.1111/j.1749-6632.1980.tb47254.x. [DOI] [PubMed] [Google Scholar]
  31. Takahashi K., Odani S., Ono T. Primary structure of rat liver Z-protein. A low-Mr cytosol protein that binds sterols, fatty acids and other small molecules. FEBS Lett. 1982 Apr 5;140(1):63–66. doi: 10.1016/0014-5793(82)80521-8. [DOI] [PubMed] [Google Scholar]
  32. Thomas K. A., Silverman R. E., Jeng I., Baglan N. C., Bradshaw R. A. Electrophoretic heterogeneity and polypeptide chain structure of the gamma-subunit of mouse submaxillary 7 S nerve growth factor. J Biol Chem. 1981 Sep 10;256(17):9147–9155. [PubMed] [Google Scholar]

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