Skip to main content
SpringerLink
Log in

Immunological comparison of the pyruvate dehydrogenase complexes from pea mitochondria and chloroplasts

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The pyruvate dehydrogenase complex (PDC) in pea (Pisum sativum L., cv. Little Marvel) was studied immunologically using antibodies to specific subunits of mammalian PDC. Pea mitochondria and chloroplasts were both found to contain PDC, but distinct differences were noted in the subunit relative molecular mass (Mr) values of the individual enzymes in the mitochondrial and chloroplast PDC complexes. In particular, the mitochondrial E3 enzyme (dihydrolipoamide dehydrogenase; EC 1.8.1.4) has a high subunit Mr value of 67 000, while the chloroplast E3 enzyme has a subunit Mr value of 52 000, similar in size to the prokaryotic, yeast ad mammalian E3 enzymes. In addition, component X (not previously noted in plant PDC) was also found to be present in two distinct forms in pea mitochondrial and chloroplast complexes. As in the case of E3, mitochondrial component X has a higher subunit Mr value (67 000) than component X from chloroplasts (48 000), which is similar in size to its mammalian counterpart. The subunit Mr value of E2 (dihydrolipoamide acetyltransferase; EC 2.3.1.12) in both mitochondria and chloroplasts (50 000) is lower than that of mammalian E2 (74 000) but similar to that of yeast E2 (58 000), and is consistent with the presence of only a single lipoyl domain. Neither mitochondria nor chloroplasts showed any appreciable cross-reactivity with antiserum to mammalian E1 (pyruvate dehydrogenase; EC 1.2.4.1). However, mitochondria cross-reacted strongly with antiserum to yeast E1, giving a single band (Mr 41 000) which is thought to be E1a. Chloroplasts showed no cross-reactivity with yeast E1, indicating that the mitochondrial E1a subunit and its chloroplast equivalent are antigenically distinct polypeptides.

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

Abbreviations

E1:

pyruvate dehydrogenase

E2:

dihydrolipoamide acetyltransferase

E3:

dihydrolipoamide dehydrogenase

Mr :

relative molecular mass

PDC:

pyruvate dehydrogenase multienzyme complex

SDS:

sodium dodecyl sulphate

References

  • Camp, P.J., Randall, D.D. (1985) Purification and characterisation of the pea chloroplast pyruvate dehydrogenase complex. Plant Physiol. 77, 571–577

    Google Scholar 

  • Day, D.A., Moore, A.L., Dry, I.B., Wiskich, J.T., Azcon-Bieto, J. (1988) Regulation of nonphosphorylating electron transport pathways in soybean cotyledon mitochondria and its implication for fat metabolism. Plant Physiol. 86, 1199–1204

    Google Scholar 

  • De Marcucci, O., Lindsay, J.G. (1985) Component X. An immunologically distinct polypeptide associated with mammalian pyruvate dehydrogenase multienzyme complex. Eur. J. Biochem. 149, 641–648

    Google Scholar 

  • De Marcucci, O.L., Hunter, A., Lindsay, J.G. (1985) Low immunogenicity of the common lipoamide dehydrogenase subunit (E3) of mammalian pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase multienzyme complexes. Biochem. J. 226, 509–517

    Google Scholar 

  • Gershwin, E., Mackay, I.R., Sturgess, A., Coppel, R.L. (1987) Identification and specificity of a cDNA encoding the 70 kd mitochondrial antigen recognized in primary biliary cirrhosis. J. Immunol. 138, 3525–3531

    Google Scholar 

  • Gopalakrishnan, S., Rahmatullah, M., Radke, G.A., Powers-Greenwood, S., Roche, T.E. (1989) Role of protein X in the function of the mammalian pyruvate dehydrogenase complex. Biochem. Biophys. Res. Commun. 160, 715–721

    Google Scholar 

  • Hodgson, J.A., De Marcucci, O.G., Lindsay, J.G. (1986) Lipoic acid is the site of substrate-dependent acetylation of component X in ox heart pyruvate dehydrogenase multienzyme complex. Eur. J. Biochem. 158, 595–600

    Google Scholar 

  • Koike, M., Koike, K. (1976) Structure, assembly and function of mammalian α-keto acid dehydrogenase complexes. Adv. Biophys. 9, 187–227

    Google Scholar 

  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685

    PubMed  Google Scholar 

  • Lawson, J.E., Behal, R.H., Reed, L.J. (1991) Disruption and mutagenesis of the Saccharomyces cerevisiae PDX1 gene encoding the protein X component of the pyruvate dehydrogenase complex. Biochemistry 30, 2834–2839

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275

    CAS  PubMed  Google Scholar 

  • Miernyk, J.A., Randall, D.D. (1987) Some properties of pea mitochondrial phospho-pyruvate dehydrogenase-phosphatase. Plant Physiol. 83, 311–315

    Google Scholar 

  • Miernyk, J.A., Camp, P.J., Randall, D.D. (1985) Regulation of plant pyruvate dehydrogenase complexes. Curr. Top. Plant Biochem. Physiol. 4, 175–190

    Google Scholar 

  • Murphy, D.J., Leech, R.M. (1977) Lipid biosynthesis from [14C]bicarbonate, [2-14C]pyruvate and (1-14C]acetate during photosynthesis by isolated spinach chloroplasts. FEBS Lett. 77, 164–168

    Google Scholar 

  • Murphy, D.J., Leech, R.M. (1978) The pathway of [14C]bicarbonate incorporation into lipids in isolated photosynthesizing spinach chloroplasts. FEBS Lett. 88, 192–196

    Google Scholar 

  • Murphy, D.J., Stumpf, P.K. (1981) The origin of chloroplastic acetyl coenzyme A. Arch. Biochem. Biophys. 212, 730–739

    Google Scholar 

  • Neagle, J.C., Lindsay, J.G. (1991) Selective proteolysis of the protein X subunit of the bovine heart pyruvate dehydrogenase complex. Biochem. J. 278, 423–427

    Google Scholar 

  • Powers-Greenwood, S.L., Rahmatullah, M., Radke, G.A., Roche, T.E. (1989) Separation of protein X from the dihydrolipoyl transacetylase component of the mammalian pyruvate dehydrogenase complex and function of protein X. J. Biol. Chem. 264, 3655–3657

    Google Scholar 

  • Randall, D.D., Rubin, P.M., Fenko, M. (1977) Plant pyruvate dehydrogenase complex purification, characterization and regulation by metabolites and phosphorylation. Biochim. Biophys. Acta 485, 336–349

    Google Scholar 

  • Randall, D.D., Williams, M., Rapp, B.J. (1981) Phosphorylation-dephosphorylation of pyruvate dehydrogenase complex from pea leaf mitochondria. Arch. Biochem. Biophys. 207, 437–444

    Google Scholar 

  • Randall, D.D., Miernyk, J.A., Fang, T.K., Budde, R.J.A., Schuller, K.A. (1990) Regulation of the pyruvate dehydrogenase complex in plants. Ann. N.Y. Acad. Sci. 573, 192–205

    Google Scholar 

  • Reed, L.J. (1974) Multienzyme complexes. Acc. Chem. Res. 7, 40–46

    Google Scholar 

  • Rice, J.E., Lindsay, J.G. (1991) Sequences directing dihydrolipoamide dehydrogenase (E3) binding are located on the 2-oxoglutarate dehydrogenase (E1) component of the mammalian 2-oxoglutarate dehydrogenase multienzyme complex. Biochem. Soc. Trans. 19, 403

    Google Scholar 

  • Rubin, P.M., Randall, D.D. (1977) Purification and characterization of pyruvate dehydrogenase complex. Arch. Biochem. Biophys. 178, 342–349

    Google Scholar 

  • Thekkumkara, T.J., Jesse, B.W., Ho, L., Raefsky, C., Pepin, R.A., Javed, A.A., Pons, G., Patel, M.S. (1987) Isolation of a cDNA clone for the dihydrolipoamide acetyltransferase component of the human liver pyruvate dehydrogenase complex. Biochem. Biophys. Res. Commun. 145, 903–907

    Google Scholar 

  • Williams, M., Randall, D.D. (1979) Pyruvate dehydrogenase complex from chloroplasts of Pisum sativum L. Plant Physiol. 64, 1099–1103

    Google Scholar 

  • Yeaman, S.J. (1989) The 2-oxo acid dehydrogenase complexes: recent advances. Biochem. J. 257, 625–632

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departments of Botany and Biochemistry, University of Glasgow, G12 8QQ, Glasgow, UK

    Anne E. Taylor, Richard J. Cogdell & J. Gordon Lindsay

Authors
  1. Anne E. Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Richard J. Cogdell
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Gordon Lindsay
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The financial support of the Agricultural and Food Research Council is gratefully acknowledged. We thank Steve Hill (Department of Botany, University of Edinburgh, UK) for advice on mitochondrial isolation, and James Neagle (Department of Biochemistry, University of Glasgow) and Ailsa Carmichael for helpful discussion.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Taylor, A.E., Cogdell, R.J. & Lindsay, J.G. Immunological comparison of the pyruvate dehydrogenase complexes from pea mitochondria and chloroplasts. Planta 188, 225–231 (1992). https://doi.org/10.1007/BF00216817

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00216817

Key words

Search

Navigation