Abstract
Isoprene synthase (ISPS) catalyzes the elimination of pyrophosphate from dimethylallyl diphosphate (DMADP) forming isoprene, a volatile hydrocarbon emitted from many plant species to the atmosphere. In the present work, immunological techniques were applied to study and localize ISPS in poplar leaves (Populus × canescens). Immunogold labeling using polyclonal antibodies generated against His-tagged recombinant ISPS protein detected ca. 44% of ISPS in the stroma of the chloroplasts and ca. 56% of gold particles attached to the stromal-facing side of the thylakoid membranes. ISPS isolated from leaves exhibited the same biochemical properties as the recombinant ISPS without the plastid-targeting peptide heterologous expressed in E. coli, whereas an additional C- or N-terminal His-tag changed the biochemical features of the recombinant enzyme with regard to temperature, pH, and substrate dependence. In comparison to the closely related class of monoterpene synthases from angiosperms and ISPS of oaks, the most striking feature of the poplar ISPS is a cooperative substrate dependence which is characteristic to enzymes with positive substrate activation. The detection of four immunoreactive bands in poplar leaf extracts with isoelectric points from 5.0 to 5.5 and a native molecular weight of ca. 51 kDa give reason for future studies on post-translational modifications of ISPS.
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Abbreviations
- DOXP/MEP:
-
1-deoxy- D-xylulose 5-phosphate/2- C-methyl-D-erythritol 4-phosphate
- DMADP:
-
Dimethylallyl diphosphate
- ISPS:
-
Isoprene synthase
- VOC:
-
Volatile organic compounds
References
Agius L, Stubbs M (2000) Investigation of the mechanism by which glucose analogues cause translocation of glucokinase in heptacytes: evidence for two glucose binding sites. Biochem J 346:413–421
Alonso WR, Croteau R (1993) Prenyltransferases and cyclases. Methods Plant Biochem 9:239–260
Bisswanger H (1994) Enzymkinetik. Theorie und Methoden 2nd edn. VCH-Wiley, Weinheim, Germany
Bohlmann J, Meyer-Gauen G, Croteau R (1998) Plant terpenoid synthases: molecular biology and phylogenetic analysis. Proc Natl Acad Sci USA 95:4126–4133
Brüggemann N, Schnitzler JP (2002a) Relationship between IDP isomerase activity and isoprene emission of oak leaves. Tree Physiol 22:1011–1018
Brüggemann N, Schnitzler JP (2002b) Diurnal variation of dimethylallyl diphosphate concentrations in oak (Quercus robur L.) leaves. Physiol Plant 115:190–196
Cardenas ML, Rabajille E, Niemeyer H (1984) Supression of kinetic cooperativity of hexokinase D (glucokinase) by competitive inhibitors. A slow transition model. Eur J Biochem 145:163–171
CRC (2003) CRC handbook of chemistry and physics, 84th edn, CRC, Boca Raton, USA
Eisenreich W, Rohdich F, Bacher A (2001) Deoxyxylulose phosphate pathway of terpenoids. Trends Plant Sci 6:78–84
Eymery F, Rey P (1999) Immunocytolocalization of CDSP 32 and CDSP 34, two chloroplastic drought-induced stress proteins in Solanum tuberosum plants. Plant Physiol Biochem 37:305–312
Fischbach RJ, Zimmer I, Steinbrecher R, Pfichner A, Schnitzler JP (2000) Monoterpene synthase activities in leaves of Picea abies (L.) Karst. and Quercus ilex L. Phytochemistry 54:257–265
Groth G, Schirwitz K (1999) Rapid purification of membrane extrinsic F1-domain of chloroplast ATP synthase in monodisperse for suitable for 3D-crystallization. Eur J Biochem 260:15–21
Guenther AB, Hewitt CN, Erickson D, Fall R, Geron C, Graedel T, Harley P, Klinger L, Lerdau M, McKay WA, Pierce T, Scholes B, Steinbrecher R, Tallamraju R, Taylor J, Zimmerman P (1995) A global model of natural volatile organic compound emissions. J Geophys Res 100:8873–8892
Kuzma J, Fall R (1993) Leaf isoprene emission rate is dependent on leaf development and the level of isoprene synthase. Plant Physiol 101:435–440
Lehning A, Zimmer I, Steinbrecher R, Brüggemann N, Schnitzler JP (1999) Isoprene synthase activity and its relation to isoprene emission in Quercus robur L. leaves. Plant Cell Environ 22:495–504
Leplé JC, Brasileiro ACM, Michel MF, Delmotte F, Jouanin L (1992) Transgenic poplars: expression of chimeric genes using four different constructs. Plant Cell Rep 11:137–141
Lichtenthaler HK (1999) The 1-deoxy-D-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Annu Rev Plant Physiol Plant Mol Biol 50:47–65
Loreto F, Velikova V (2001) Isoprene produced by leaves protects the photosynthetic apparatus against ozone damage, quenches ozone products, and reduces lipid peroxidation of cellular membranes. Plant Physiol 127:1781–1787
Loreto F, Mannozzi M, Maris C, Nascetti P, Ferranti F, Pasqualini S (2001) Ozone quenching properties of isoprene and its antioxidant role in leaves. Plant Physiol 126:993–1000
Mayrhofer S, Heizmann U, Magel E, Eiblmeier M, Müller A, Rennenberg H, Hampp R, Schnitzler J-P, Kreuzwieser J (2004) Carbon balance in the leaves of young poplar trees. Plant Biol 6:730–745
Miller B, Oschinski C, Zimmer W (2001) First isolation of an isoprene synthase gene from poplar and successful expression of the gene in Escherichia coli. Planta 213:483–487
Monson RK, Jaeger CH, Adams III WW, Driggers EM, Silver GM, Fall R (1992) Relationship among isoprene emission rate, photosynthesis, and isoprene synthase activity as influenced by temperature. Plant Physiol 98:1175–1180
Parmryd I, Shipton CA, Swiezewska E, Dallner G, Andersson B (1997) Chloroplastic prenylated proteins. FEBS Lett 414:527–531
Parmryd I, Andersson B, Dallner G (1999) Protein prenylation in spinach chloroplasts. Proc Natl Acad Sci USA 96:10074–10079
Rasmussen RA (1970) Isoprene: identified as a forest-type emission to the atmosphere. Environ Sci Technol 4:667–671
Rosenstiel TN, Potosnak MJ, Griffin KL, Fall R, Monson RK (2003) Increased CO2 uncouples growth from isoprene emission in an agriforest ecosystem. Nature 421:256–259
Rübsamen H, Khandker R, Witzel H (1974) Sigmoidal kinetics of a monomeric ribonuclease I due to ligand-induced shifts of conformation equilibrium. Hoppe-Seyler’s Zeit f Physiol Chem 355:687–708
Sanadze GA (1991) Isoprene effect light-dependent emission of isoprene by green parts of plants. In: Sanadze GA (ed) Trace gas emissions by plants. Academic, San Diego, USA, pp 135–152
Schnitzler JP, Arenz R, Steinbrecher R, Lehning A (1996) Characterization of an isoprene synthase from leaves of Quercus petraea (Mattuschka) Liebl. Bot Acta 109:216–221
Schnitzler JP, Jungblut TP, Feicht C, Köfferlein M, Langebartels C, Heller W, Sandermann Jr H (1997) UV-B induction of flavonoid biosynthesis in Scots pine (Pinus sylvestris L.) seedlings. Trees 11:162–168
Schnitzler JP, Steinbrecher R, Zimmer I, Steigner D, Fladung M (2004) Hybridisation of European oaks (Quercus ilex × Q. robur) results in a mixed isoprenoid emitter type. Plant Cell Environ 27:585–594
Scholefield PA, Kieron J, Doick KJ, Herbert B, Hewitt CN, Schnitzler JP, Pinelli P, Loreto F (2004) Impact of rising CO2 on VOC emissions: isoprene emission from Phragmites australis growing at elevated CO2 in a natural carbon dioxide spring. Plant Cell Environ 27:381–392
Sharkey TD, Yeh S (2001) Isoprene emission from plants. Annu Rev Plant Phys Plant Mol Biol 52:407–436
Silver GM, Fall R (1991) Enzymatic synthesis of isoprene from dimethylallyl diphosphate in aspen leaf extracts. Plant Physiol 97:1588–1591
Silver GM, Fall R (1995) Characterization of aspen isoprene synthase, an enzyme responsible for leaf isoprene emission to the atmosphere. J Biol Chem 270:13010–13016
Smith MD, Licatalosi DD, Thompson JE (2000) Co-association of cytochrome f catabolites plastid-lipid-associated protein with chloroplast lipid particles. Plant Physiol 124:211–221
Turner GT, Gershenzon J, Nielson EE, Froehlich JE, Croteau R (1999) Limonene synthase, the enzyme responsible for monoterpene biosynthesis in peppermint, is localized to leucoplasts of oil gland secretory cells. Plant Physiol 120:879–886
Wildermuth MC, Fall R (1996) Light-dependent isoprene emission. Characterization of a thylakoid-bound isoprene synthase in Salix discolor chloroplasts. Plant Physiol 112:171–182
Wildermuth MC, Fall R (1998) Biochemical characterization of stromal and thylakoid-bound isoforms of isoprene synthase in willow leaves. Plant Physiol 116:1111–1123
Wolfertz M, Sharkey TD, Boland W, Kühnemann F (2004) Rapid regulation of the methylerythritol 4-pathway during isoprene synthesis. Plant Physiol 135:1939–1945
Xu LZ, Harrison RW, Weber IT, Pilkis SJ (1995) Human ß-cell glucokinase. J Biol Chem 270:9939–9946
Acknowledgements
The authors thank T. Fenning (Max-Planck-Institute for Chemical Ecology, Jena, Germany) and B. Miller (University of Freiburg, Germany) for critical reading of the manuscript and helpful comments. In addition, we acknowledge the help of W. Heller (GSF Research Center, Neuherberg, Germany) who enabled the production of the antibody and A. Vieler (Technical University of Munich, Germany) for her technical assistance with the immunogold labeling. The work was financially supported by the German Federal Ministry of Education and Research (BMBF) in the framework of the national joint research project ‘AFO2000’ (Atmosphären-Forschungsprogramm 2000) and the German Research Foundation (DFG) in the framework of the research project ‘Poplar—a model to address tree-specific questions’.
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Schnitzler, JP., Zimmer, I., Bachl, A. et al. Biochemical properties of isoprene synthase in poplar (Populus × canescens). Planta 222, 777–786 (2005). https://doi.org/10.1007/s00425-005-0022-1
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DOI: https://doi.org/10.1007/s00425-005-0022-1