Skip to main content
SpringerLink
Log in

Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Acyl-CoA binding proteins (ACBPs) are small (ca. 10 kDa) highly-conserved cytosolic proteins that bind long-chain acyl-CoAs. A novel cDNA encoding ACBP1, a predicted membrane protein of 24.1 kDa with an acyl-CoA binding protein domain at its carboxy terminus, was cloned from Arabidopsis thaliana. At this domain, ACBP1 showed 47% amino acid identity to Brassica ACBP and 35% to 40% amino acid identity to yeast, Drosophila, bovine and human ACBPs. Recombinant (His)6-ACBP1 fusion protein was expressed in Escherichia coli and was shown to bind 14[C]oleoyl-CoA. A hydrophobic domain, absent in the 10 kDa ACBPs, was located at the amino terminus of ACBP1. Using antipeptide polyclonal antibodies in western blot analysis, ACBP1 was shown to be a membrane-associated glycosylated protein with an apparent molecular mass of 33 kDa. The ACBP1 protein was also shown to accumulate predominantly in siliques and was localized to the seed within the silique. These results suggest that the biological role of ACBP1 is related to lipid metabolism in the seed, presumably in which acyl-CoA esters are involved. Northern blot analysis showed that the 1.4 kb ACBP1 mRNA was expressed in silique, root, stem, leaf and flower. Results from Southern blot analysis of genomic DNA suggest the presence of at least two genes encoding ACBPs in Arabidopsis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Alho H, Costa E, Ferrero P, Fujimoto M, Cosenza-Murphy D, Guidotti A: Diazepam-binding inhibitor: a neuropeptide located in selected neuronal populations of rat brain. Science 229: 179–182 (1985).

    Google Scholar 

  2. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: Current Protocols in Molecular Biology. Greene Publishing Associates/Wiley-Interscience, New York (1995).

    Google Scholar 

  3. Bradford MM: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  4. Browse J: Glycerolipid synthesis. Annu Rev Plant Physiol Plant Mol Biol 42: 467–506 (1991).

    Google Scholar 

  5. Chye M-L, Cheung K-Y, Xu J: Characterization of TSCL, a nonviral retroposon from Arabidopsis thaliana. Plant Mol Biol 35: 893–903 (1997).

    Google Scholar 

  6. Engeseth NJ, Pacovsky RS, Newman T, Ohlrogge JB: Characterization of an acyl-CoA-binding protein from Arabidopsis thaliana. Arch Biochem Biophys 331: 55–62 (1996).

    Google Scholar 

  7. Gray PW, Glaister D, Seeburg PH, Guidotti A, Costa E: Cloning and expression of cDNA for human diazepam binding inhibitor, a natural ligand of an allosteric regulatory site of the γ-aminobutyric acid type A receptor. Proc Natl Acad Sci USA 83: 7547–7551 (1986).

    Google Scholar 

  8. Guidotti A, Forchetti CM, Corda MG, Konkel D, Bennett CD, Costa E: Isolation, characterization and purification to homogeneity of an endogenous polypeptide with agonistic action on benzodiazepine receptors. Proc Natl Acad Sci USA 80: 3531–3535 (1983).

    Google Scholar 

  9. Harwood JL: Plant acyl lipids: structure, distribution and analysis. In: Stumpf PK, Conn EE (eds) The Biochemistry of Plants: A Comprehensive Treatise, vol. 4, pp. 1–55. Academic Press, New York (1990).

    Google Scholar 

  10. Hills MJ, Dann R, Lydiate D, Sharpe A: Molecular cloning of a cDNA from Brassica napus L. for a homologue of acyl-CoA binding protein. Plant Mol Biol 25: 917–920 (1994).

    Google Scholar 

  11. Hong Y, Takano M, Liu C-M, Gasch A, Chye M-L, Chua N-H: Expression of three members of the calcium-dependent protein kinase gene family in Arabidopsis thaliana. Plant Mol Biol 30: 1259–1275 (1996).

    Google Scholar 

  12. Kader J-P: Lipid-transfer proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 47: 627–654 (1996).

    Google Scholar 

  13. Klein P, Kanehisa M, De Lisi C: The detection and classification of membrane-spanning proteins. Biochim Biophys Acta 815: 468–476 (1985).

    Google Scholar 

  14. Kolmer M, Alho H, Costa E, Pani L: Cloning and tissuespecific functional characterization of the promoter of the rat diazepam binding protein inhibitor, a peptide with multiple biological actions. Proc Natl Acad Sci USA 90: 8439–8443 (1993).

    Google Scholar 

  15. Kolmer M, Roos C, Tirronen M, Myohanen S, Alho H: Tissue-specific expression of the diazepam-binding inhibitor in Drosophila melanosgaster: cloning, structure and localization of the gene. Mol Cell Biol 14: 6983–6995 (1994).

    Google Scholar 

  16. Kragelund BB, Andersen KV, Madsen JC, Knudsen J and Poulsen FM: Three-dimensional structure of the complex between acyl-coenzyme A binding protein and palmitoylcoenzyme A. J Mol Biol 230: 1260–1277 (1993).

    Google Scholar 

  17. Kyte J, Doolittle RF: A simple method of displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).

    Google Scholar 

  18. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).

    Google Scholar 

  19. Lemieux B, Koornneef M, Feldmann KA: Epicuticular wax and eceriferum mutants. In: Meyerowitz EM, Somerville CR (eds) Arabidopsis, pp. 1031–1047. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1994).

    Google Scholar 

  20. Lihrmann I, Plaquevent J-C, Tostivint H, Raijmakers R, Tonon M-C, Conlon J-M, Vaudry H: Frog diazepam-binding inhibitor: peptide sequence, cDNA cloning, and expression in the brain. Proc Natl Acad Sci USA 91: 6899–6903 (1994).

    Google Scholar 

  21. Lutcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  22. Mandrup S, Hummel R, Ravn S, Jensen G, Andreasen PH, Gregersen N, Knudsen J, Kristiansen K: Acyl-CoA binding protein/diazepam binding inhibitor gene and pseudogenes. A typical housekeeping gene family. J Mol Biol 228: 1011–1022 (1992).

    Google Scholar 

  23. Marquardt H, Todaro GJ, Shoyab M: Complete amino acid sequences of bovine and human endozepines. Homology with rat diazepam binding inhibitor. J Biol Chem 261: 9727–9731 (1986).

    Google Scholar 

  24. Mikkelsen J, Hojrup P, Nielsen PF, Roepstorff P, Knudsen J: Amino acid sequence of acyl-CoA-binding sequence from cow liver. Biochem J 245: 857–861 (1987).

    Google Scholar 

  25. Mikkelsen J, Knudsen J: Acyl-CoA binding protein from cow. Biochem J 248: 709–714 (1987).

    Google Scholar 

  26. Mocchetti E, Einstein R, Brosius J: Putative diazepam binding inhibitor peptide: cDNA clones from rat. Proc Natl Acad Sci USA 83: 7221–7225 (1986).

    Google Scholar 

  27. Nagy F, Kay SA, Chua N-H: Analysis of gene expression in transgenic plants. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, pp. B4: 1–29. Kluwer Academic Publishers, Dordrecht, Netherlands (1988).

    Google Scholar 

  28. Ohlrogge JB, Kuhn DN, Stumpf PK: Subcellular localization of acyl carrier protein in leaf protoplasts of Spinacia oleracea. Proc Natl Acad Sci USA 76: 1194–1198 (1979).

    Google Scholar 

  29. Rasmussen JT, Borchers T, Knudsen J: Comparison of the binding affinities of acyl-CoA binding protein and fatty acid binding protein for long-chain acyl-CoA esters. Biochem J 265: 849–855 (1990).

    Google Scholar 

  30. Rasmussen JT, Faergeman NJ, Kristiansen K, Knudsen J: Acyl-CoA binding protein (ACBP) can mediate intermembrane acyl-CoA transport and donate acyl-CoA for betaoxidation and glycerolipid synthesis. Biochem J 299: 165–170 (1994).

    Google Scholar 

  31. Rasmussen JT, Rosendal J, Knudsen J: Interaction of acyl-CoA binding protein (ACBP) on processes for which acyl-CoA is a substrate, product or inhibitor. Biochem J 292: 907–913 (1993).

    Google Scholar 

  32. Reddy AS, Ranganathan B, Haisler RM, Swize MA: A cDNA encoding acyl-CoA-binding protein from cotton (accession No. U35015). (PGR96-028). Plant Physiol 111: 348 (1996).

    Google Scholar 

  33. Rose TM, Schultz ER, Todaro GJ: Molecular cloning of the gene for the yeast homolog (ACB) of diazepam binding inhibitor/endozepine/acyl-CoA-binding protein. Proc Natl Acad Sci USA 89: 1287–1291 (1992).

    Google Scholar 

  34. Roughan PG, Holland R, Slack CR: On the control of longchain fatty acid synthesis in isolated intact spinach (Spinacia oleracea) chloroplasts. Biochem J 184: 193–202 (1979).

    Google Scholar 

  35. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  36. Sanger F, Nicklen S, Coulsen AR: DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  37. Shoyab M, Gentry LE, Marquardt H, Todaro GJ: Isolation and characterization of a putative endogenous benzodiazepineoid (endozepine) from bovine and human brain. J Biol Chem 261: 11968–11973 (1986).

    Google Scholar 

  38. Slabas AR, Fawcett T: The biochemistry and molecular biology of plant lipid biosynthesis. Plant Mol Biol 19: 169–191 (1992).

    Google Scholar 

  39. Smith JA, Krauss MR, Borkird C, Sung ZR: A nuclear protein associated with cell divisions in plants. Planta 174: 462–472 (1988).

    Google Scholar 

  40. Studier FM, Moffat BA Use of bacteriophage T7 RNA polymerase to direct selective high-expression of cloned genes. J Mol Biol 189: 113–130 (1986).

    Google Scholar 

  41. Swinnen JV, Esquenet M, Rossels J, Claessens F, Rombauts W, Heyns W, Verhoeven G: A human gene encoding diazepam-binding inhibitor/acyl-CoA-binding protein: transcription and hormonal regulation in the androgen-sensitive human prostatic adenocarcinoma cell line LNCaP. DNA Cell Biol 15: 197–208 (1996).

    Google Scholar 

  42. Takano E, Maki M, Mori H, Hatanaka M, Marti T, Titani K, Kannagi R, Ooi T, Murachi T: Pig heart calpastatin: identi-fication of repetitive domain structure and anamalous behaviour in polyacylamide gel electrophoresis. Biochemistry 27: 1964–1972 (1988).

    Google Scholar 

  43. Todaro GJ, Rose TM, Shoyab M: Human DBI (endozepine): relationship to a homologous membrane associated protein (MA-DBI). Neuropharmacology 30: 1373–1380 (1991).

    Google Scholar 

  44. Webb NR, Rose TM, Malik N, Marquardt H, Shoyab M, Todaro GJ, Lee DC: Bovine and human cDNA sequences encoding a putative benzodiazepine receptor ligand. DNA 6: 71–79 (1987).

    Google Scholar 

  45. Weber K, Pringle JR, Osborne M: Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Meth Enzymol 91: 263–277 (1971).

    Google Scholar 

  46. Yanisch-Perron C, Vieira J, Messing J: Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103–119 (1985).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany, University of Hong Kong, Pokfulam Road, Hong Kong, China

    Mee-Len Chye

Authors
  1. Mee-Len Chye
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chye, ML. Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain. Plant Mol Biol 38, 827–838 (1998). https://doi.org/10.1023/A:1006052108468

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006052108468

Search

Navigation