Abstract
We have purified for the first time from green leaves a multifunctional protein (MFP) involved in fatty acid β-oxidation. The protein, designated MFP IV, was extracted from green leaves of three-week-old cucumber (Cucumis sativus L.) plants. Chromatography on cation exchanger, separation on hydroxylapatite, and fast-protein liquid chromatography on Phenylsuperose led to a more than 7000-fold purification and to the isolation of an apparently homogeneous 80-kDa monomeric protein. This protein is immunologically related to the glyoxysomal MFP II, as evidenced by immunodecoration with antiserum raised against MFP II. Comparison of molecular masses of all MFPs presently known revealed that the MFP prepared from green leaves (MFP IV) is distinct from MFP II (76.5 kDa) and MFP I (74 kDa) from dark-grown cotyledons. By including other properties in this comparison, we demonstrated that MFP IV can also be distinguished from the glyoxysomal MFP III (81 kDa) and the bacterially expressed MFP-a (80 kDa). Moreover, MFP IV is a constituent of leaf peroxisomes and contains the activities of 2-enoyl-CoA hydratase (EC 4.2.1.17),l-3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) and 3-hydroxyacyl-CoA epimerase.
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Abbreviations
- MFP:
-
multifunctional protein
References
Behrends, W., Engeland, K., Kindl, H. (1988) Characterization of two forms of the multifunctional protein acting in the glyoxysomal fatty acid β-oxidation. Arch. Biochem. Biophys.263, 161–169
Binstock, J.F., Schulz, H. (1981) Fatty acid oxidation complex fromEscherichia coli. Methods Enzymol.71, 403–411
Birkhan, R., Kindl, H. (1990) Re-activation of the expression of glyoxysomal genes in green plant tissue. Z. Naturforsch.45C, 68–72
Cooper, T.G., Beevers, H. (1969) Mitochondria and glyoxysomes from castor bean endosperm. J. Biol. Chem.244, 3507–3513
De Bellis, L., Nishimura, M. (1991) Development of enzymes of the glyoxylate cycle during senescence of pumpkin cotyledons. Plant Cell Physiol.32, 555–561
Engeland, K., Kindl, H. (1991a) Evidence for a peroxisomal fatty acid β-oxidation involving D-3-hydroxyacyl-CoAs. Eur. J. Biochem.200, 1717–1718
Engeland, K., Kindl, H. (1991b) Purification and characterization of a plant peroxisomal 3-cis-enoyl-CoA and 3-trans-enoyl-CoA. Eur. J. Biochem.196, 699–705
Frevert, J., Kindl, H. (1980a) A bifunctional enzyme from glyoxysomes: Purification of a protein possessing enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activity. Eur. J. Biochem.107, 79–86
Frevert, J., Kindl, H. (1980b) Purification of glyoxysomal acetyl- CoA acyltransferase. Hoppe-Seyler's Z. Physiol. Chem.361, 537–542
Gerbling, H., Gerhardt, B. (1987) Activation of fatty acids by non-glyoxysomal peroxisomes. Planta171, 386–392
Gerhardt, B. (1985) Substrate specificity of peroxisomal acyl-CoA oxidase. Phytochemistry24, 351–352
Gerhardt, B. (1987) Peroxisomes and fatty acid degradation. Methods Enzymol.148, 517–525
Gerhardt, B. (1992) Fatty acid degradation in plants. Progr. Lipid Res.31, 417–46
Goldman, P., Vageolos, P.R. (1961) The specificity of triglyceride synthesis from diglycerides in chicken adipose tissue. J. Biol. Chem.236, 2620–2623
Gut, H., Matile, P. (1988) Apparent induction of key enzymes of the glyoxylate cycle in senescent barley leaves. Planta176, 548–550
Huang, A.H.C. (1992) Oil bodies and oleosins in seeds. Annu. Rev. Plant Physiol.43, 177–200
Kindl, H. (1982) Glyoxysome biogenesis via cytosolic pools in cucumber. Ann. N. Y. Acad. Sci.386, 314–328
Kindl, H. (1987) β-Oxidation of fatty acids by specific organelles. In: The biochemistry of plants, Vol. 9, pp. 31–52, Stumpf, P.K., Conn, E.E., eds. Academic Press, London
Kindl, H. (1993) Fatty acid degradation in plant peroxisomes-function and biosynthesis of the enzymes involved. Biochimie75, 225–230
Kirsch, T., Löffler, H.-G., Kindl, H. (1986) Plant acyl-CoA oxidase. Purification, characterization and monomeric apoprotein. J. Biol. Chem.26, 8570–8575
Kunau, W.H., Kionka, C., Ledebur, A., Mateblowski, M., Moreno de la Garza, M., Schultz-Borchard, U., Thieringer, R., Veenhuis, M. (1987) β-Oxidation systems in eukaryotic microorganisms. In: Peroxisomes in biology in medicine, pp. 128–140, Fahimi, H.D., Sies, H., eds. Springer, Berlin
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227, 680–685
Murphy, D.J. (1990) Storage lipid bodies in plants and other organisms. Progr. Lipid Res.29, 299–324
Murphy, D.J. (1993) Structure, function and biogenesis of storage lipid bodies and oleosins in plants. Progr. Lipid Res.32, 247–280
Preisig-Müller, R., Gühnemann-Schäfer, K., Kindl, H. (1994) Domains of the tetrafunctional protein acting in glyoxysomal fattyacid β-oxidation. Demonstration of epimerase and isomerase activities on a peptide lacking hydratase activity. J. Biol. Chem.269, 20475–20481
Schiefer, S., Teifel, W., Kindl, H. (1976) Plant microbody proteins. I. Purification and characterization of catalase from leaves ofLens culinaris. Hoppe-Seyler's Z. Physiol. Chem.357, 177–186
Schnarrenberger, C., Oeser, A., Tolbert, N.E. (1972) Isolation of plastids from sunflower cotyledons during germination. Plant Physiol.50, 55–59
Schulz, H. (1991) Beta oxidation of fatty acids. Biochim. Biophys. Acta1081, 109–120
Schulz, H., Kunau, W.H. (1987) Beta-oxidation of unsaturated fatty acids. A revised pathway. Trends Biochem. Sci.12, 403–406
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This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie.
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Gühnemann-Schäfer, K., Kindl, H. The leaf peroxisomal form (MFP IV) of multifunctional protein functioning in fatty-acid β-oxidation. Planta 196, 642–646 (1995). https://doi.org/10.1007/BF01106755
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DOI: https://doi.org/10.1007/BF01106755