Radioimmunoassay and Western Blot Analysis of Acyl Carrier Protein Isoforms in Plants

  • T. M. Kuo
  • J. B. Ohlrogge
Part of the Modern Methods of Plant Analysis book series (MOLMETHPLANT, volume 4)


Acyl carrier protein (ACP) is the central cofactor protein to which acyl chains are covalently linked as thioesters during de novo fatty acid synthesis and subsequent elongation and desaturation reactions in plants (Stumpf 1980). In yeast and animals, ACP is an integral part of the multifunctional polypeptide, fatty acid synthase, but in bacteria and plants, ACP is a separate, small polypeptide (ca. 9 kDa MW) which is acidic and heat stable (Majerus and Vagelos 1967; Stumpf 1980; Wakil et al. 1983). Despite this basic organizational difference, the structure of ACP has been highly conserved during evolution as evidenced by greater than 25% homology among the primary sequences of an animal ACP domain, bacterial and plant ACPs (Kuo and Ohlrogge 1984a), and by the interchangeability of bacterial and plant ACPs in several fatty acid biosynthetic reactions (Simoni et al. 1967).


Fatty Acid Synthesis Castor Bean Acyl Carrier Protein Polytron Homogenizer Antibody Binding Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



acyl carrier protein




2-(N-morpholino)ethanesulfonic acid


high-performance liquid chromatography






bovine serum albumin


phosphate-buffered saline


polyethylene glycol


days after flowering


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  1. Benade L, Ihle JN (1982) Principles of radioimmunoassays and related techniques. In: Marchalonis JJ, and Wan GW (eds) Antibody as a tool. Wiley, New York, pp 163–187Google Scholar
  2. Bolton AE, Hunter WM (1973) The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent-application to the radioimmunoassay. Biochem J 133: 529–539PubMedGoogle Scholar
  3. Chafouleas JG, Dedman JR, Means AR (1982) Radioimmunoassay of calmodulin. Methods Enzymol 84: 138–147PubMedCrossRefGoogle Scholar
  4. Creighton WD, Lambert PH, Miescher PA (1973) Detection of antibodies and soluble antigen-antibody complexes by precipitation with polyethylene glycol. J Immunol 111: 1219–1227PubMedGoogle Scholar
  5. Hawker CD (1973) Radioimmunoassay and related methods. Anal Chem 45: 878A - 890ACrossRefGoogle Scholar
  6. Hej PB, Svendsen I (1984) Barley chloroplasts contain two acyl carrier proteins coded for by different genes. Carlsberg Res Commun 49: 483–492CrossRefGoogle Scholar
  7. Hunter WM, Greenwood FC (1962) Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature 194: 495–496PubMedCrossRefGoogle Scholar
  8. Johnson DA, Gautsch JW, Sportsman JH, Elder JH (1984) Improved technique utilising nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1: 3–8CrossRefGoogle Scholar
  9. Kuo TM, Ohlrogge JB (1984a) The primary structure of spinach acyl carrier protein. Arch Biochem Biophys 234: 290–296PubMedCrossRefGoogle Scholar
  10. Kuo TM, Ohlrogge JB (1984b) A novel, general radioimmunoassay for acyl carrier proteins. Anal Biochem 136: 497–502PubMedCrossRefGoogle Scholar
  11. Kuo TM, Ohlrogge JB (1984c) Acylation of plant acyl carrier proteins by acyl-acyl carrier protein synthetase from Escherichia coli. Arch Biochem Biophys 230: 110–116PubMedCrossRefGoogle Scholar
  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bac-teriophage T4. Nature 227: 680–685PubMedCrossRefGoogle Scholar
  13. Majerus PW, Vagelos PR (1967) Fatty acid biosynthesis and the role of the acyl carrier protein. In: Paoletti R, Kritchevsky D (eds) Advances in lipid research, vol 5. Academic Press, New York, pp 1–33Google Scholar
  14. Nakatani HY, Barber J (1977) An improved method for isolating chloroplasts retaining their outer membranes. Biochim Biophys Acta 461: 510–512CrossRefGoogle Scholar
  15. Ohlrogge JB, Kuo TM (1984) Control of lipid synthesis during soybean seed development: enzymatic and immunochemical assay of acyl carrier protein. Plant Physiol (Bethesda) 74: 622–625CrossRefGoogle Scholar
  16. Ohlrogge JB, Kuo TM (1985) Plants have isoforms for acyl carrier protein that are expressed differently in different tissues. J Biol Chem 260: 8032–8037PubMedGoogle Scholar
  17. Ohlrogge JB, Kuhn DN, Stumpf PK (1979) Subcellular localization of acyl carrier protein in leaf protoplasts of Spinacia oleracea. Proc Natl Acad Sci USA 76: 1194–1198PubMedCrossRefGoogle Scholar
  18. Reichlin M (1980) Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods Enzymol 70: 159–165PubMedCrossRefGoogle Scholar
  19. Rice RH, Means GE (1971) Radioactive labeling of proteins in vitro. J Biol Chem 246: 831–832PubMedGoogle Scholar
  20. Rock CO, Cronan JE Jr (1979) Solubilization, purification, and salt activation of acyl-acyl carrier protein synthetase from Escherichia coli. J Biol Chem 254: 7116–7122PubMedGoogle Scholar
  21. Rock CO, Cronan JE Jr (1980) Improved purification of acyl camer protein. Anal Biochem 102: 362–364PubMedCrossRefGoogle Scholar
  22. Rock CO, Garwin JL (1979) Preparative enzymatic synthesis and hydrophobic chromatography of acyl-acyl carrier protein. J Biol Chem 254: 7123–7128PubMedGoogle Scholar
  23. Simoni RD, Criddle RS, Stumpf PK (1967) Fat metabolism in higher plants. XXXI. Purification and properties of plant and bacterial acyl carrier proteins. J Biol Chem 242: 573–581PubMedGoogle Scholar
  24. Stumpf PK (1980) Biosynthesis of saturated and unsaturated fatty acids. In: Stumpf PK (ed) The Biochemistry of plants, vol 4. Academic Press, New York, pp 177–204Google Scholar
  25. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Nad Acad Sci USA 76: 4350–4354CrossRefGoogle Scholar
  26. Tzeng M-C (1983) A sensitive, rapid method for monitoring sodium dodecyl sulfate-poly-acrylamide gel electrophoresis by chromophoric labeling. Anal Biochem 128: 412–414PubMedCrossRefGoogle Scholar
  27. Wakil SJ, Stoops JK, Joshi VC (1983) Fatty acid synthesis and its regulation. Annu Rev Biochem 52: 537–579PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1986

Authors and Affiliations

  • T. M. Kuo
  • J. B. Ohlrogge

There are no affiliations available

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