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Licensed Unlicensed Requires Authentication Published by De Gruyter March 19, 2014

Lanthanides as substitutes for calcium ions in the activation of plant α-type phospholipase D

  • Lars Dressler , Ralph Golbik and Renate Ulbrich-Hofmann EMAIL logo
From the journal Biological Chemistry

Abstract

Most types of phospholipase D (PLD) from plants contain a C2 domain and are activated by Ca2+ ions. In this study, other metal ions such as Mg2+, La3+, Ce3+, Tb3+ and Y3+ were examined as effectors of recombinantly produced α-type PLD from white cabbage. All the rare earth ions were able to substitute for Ca2+. The activation curves and displacement experiments reflect a 10- to 50-fold higher affinity of PLD for these ions than for Ca2+; however, the maximum activity attained only 36% of that in the presence of Ca2+. Mg2+ displaced Ca2+ without being able to activate PLD. All ions were bound to the substrate micelles consisting of phosphatidyl-p-nitrophenol, Triton X-100 and SDS (1:8:1, by mole). The affinity of rare earth ions to the micelles was 100-fold higher than that of Ca2+ and Mg2+. A conformational change of the enzyme induced by the low affinity but specific binding of Ca2+ ions is concluded to be essential for maximal PLD activity. As demonstrated by the measurement of Tb3+ fluorescence, the substitution of Ca2+ by rare earth ions provides a new avenue for studying the enigmatic role of Ca2+ ions in the modulation of PLD activity in plants.


Corresponding author: Renate Ulbrich-Hofmann, Institute of Biochemistry and Biotechnology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, D-06120 Halle, Germany, e-mail:

Acknowledgments

We thank the group of Alfred Blume, Institute of Chemistry, Martin-Luther University Halle-Wittenberg, for the support in DLS measurements. Moreover, we are thankful to Gary Sawers, Institute of Biology, Martin-Luther University Halle-Wittenberg, for reading the manuscript. The financial support by the Graduiertenkolleg 1026 (Deutsche Forschungsgemeinschaft, Bonn, Germany) is gratefully acknowledged.

References

Abdelkafi, S. and Abousalham, A. (2011). Kinetic study of sunflower phospholipase Dα: Interactions with micellar substrate, detergent and metals. Plant Physiol. Biochem. 49, 752–757.10.1016/j.plaphy.2011.02.002Search in Google Scholar

Chaudhuri, D., Horrocks, W.DeW., Jr., Amburgey, J.C., and Weber, D.J. (1997). Characterization of lanthanide ion binding to the EF-hand protein S100β by luminescence spectroscopy. Biochemistry 36, 9674–9680.10.1021/bi9704358Search in Google Scholar

Cho, W. and Stahelin, R.V. (2006). Membrane binding and subcellular targeting of C2 domains. Biochim. Biophys. Acta 1761, 838–849.10.1016/j.bbalip.2006.06.014Search in Google Scholar

D’Arrigo, P., Piergianni, V., Scarcelli, D., and Servi, S. (1995). A spectrophotometric assay for phospholipase D. Anal. Chim. Acta 304, 249–254.10.1016/0003-2670(94)00613-QSearch in Google Scholar

Damnjanovic, J. and Iwasaki, Y. (2013). Phospholipase D as a catalyst: application in phospholipid synthesis, molecular structure and protein engineering. J. Biosci. Bioeng. 116, 271–280.10.1016/j.jbiosc.2013.03.008Search in Google Scholar PubMed

Dippe, M., Dressler, L., and Ulbrich-Hofmann, R. (2014). Fe(III)-resorcylate as a spectrophotometric probe for phospholipid-cation interactions. Anal. Biochem. 445, 54–59.10.1016/j.ab.2013.10.008Search in Google Scholar PubMed

Gottlin, E.B., Rudolph, A.E., Zhao, Y., Matthews, H.R., and Dixon, J.E. (1998). Catalytic mechanism of the phospholipase D superfamily proceeds via a covalent phosphohistidine intermediate. Proc. Natl. Acad. Sci. USA 95, 9202–9207.10.1073/pnas.95.16.9202Search in Google Scholar PubMed PubMed Central

Iwasaki, Y., Nakano, H., and Yamane, T. (1994). Phospholipase D from Streptomyces antibioticus: cloning, sequencing, expression, and relationship to other phospholipases. Appl. Microbiol. Biotechnol. 42, 290–299.10.1007/s002530050252Search in Google Scholar

Jang, J.-H., Lee, C.S., Hwang, D., and Ryu, S.H. (2012). Understanding of the roles of phospholipase D and phosphatidic acid through their binding partners. Progr. Lipid Res. 51, 71–81.10.1016/j.plipres.2011.12.003Search in Google Scholar PubMed

Kolesnikov, Y.S., Nokhrina, K.P., Kretynin, S.V., Volotovski, I.D., Martinec, J., Romanov, G.A., and Kravets, V.S. (2012). Molecular structure of phospholipase D and regulatory mechanisms of its activity in plant and animal cells. Biochemistry (Moscow) 77, 1–14.10.1134/S0006297912010014Search in Google Scholar PubMed

Leiros, I., Secundo, F., Zambonelli, C., Servi, S., and Hough, E. (2000). The first crystal structure of a phospholipase D. Structure 8, 655–667.10.1016/S0969-2126(00)00150-7Search in Google Scholar

Lerchner, A., Mansfeld, J., Kuppe, K., and Ulbrich-Hofmann, R. (2006). Probing conserved amino acids in phospholipase D (Brassica oleracea var. capiata) for their importance in hydrolysis and transphosphatidylation activity. Protein Eng. Des. Sel. 19, 443–452.10.1093/protein/gzl028Search in Google Scholar

McDermott, M., Wakelam, M.J.O., and Morris, A.J. (2004). Phospholipase D. Biochem. Cell Biol. 82, 225–253.10.1139/o03-079Search in Google Scholar

Pappan, K., Zheng, L., Krishnamoorthi, R., and Wang, X. (2004). Evidence for and characterization of Ca2+ binding to the catalytic region of Arabidopsis thaliana phospholipase Dβ. J. Biol. Chem. 279, 47833–47839.10.1074/jbc.M402789200Search in Google Scholar

Petersheim, M., Halladay, H.N., and Blodnieks, J. (1989). Tb3+ and Ca2+ binding to phosphatidylcholine. A study comparing data from optical, NMR, and infrared spectroscopies. Biophys. J. 56, 551–557.10.1016/S0006-3495(89)82702-XSearch in Google Scholar

Pidcock, E. and Moore, G.R. (2001). Structural characteristics of protein binding sites for calcium and lanthanide ions. J. Biol. Inorg. Chem. 6, 479–489.10.1007/s007750100214Search in Google Scholar PubMed

Pintacuda, G., John, M., Su, X.-C., and Otting, G. (2007). NMR structure determination of protein-ligand complexes by lanthanide labeling. Acc. Chem. Res. 40, 206–212.10.1021/ar050087zSearch in Google Scholar PubMed

Ponting, C.P. and Kerr, I.D. (1996). A novel family of phospholipase D homologues that includes phospholipid synthases and putative endonucleases: identification of duplicated repeats and potential active site residues. Protein Sci. 5, 914–922.10.1002/pro.5560050513Search in Google Scholar PubMed PubMed Central

Qin, C. and Wang, X. (2002). The Arabidopsis D family. Characterization of a calcium-independent and phosphatidylcholine-selective PLDζ1 with distinct regulatory domains. Plant Physiol. 128, 1057–1068.10.1104/pp.010928Search in Google Scholar PubMed PubMed Central

Qin, W., Pappan, K., and Wang, X. (1997). Molecular heterogeneity of phospholipase D (PLD). Cloning of PLDγ and regulation of plant PLDγ, -β, and -α by polyphosphoinositides and calcium. J. Biol. Chem. 272, 28267–28273.10.1074/jbc.272.45.28267Search in Google Scholar PubMed

Reich, S., Golbik, R.P., Geissler, R., Lilie, H., and Behrens, S.-E. (2010). Mechanisms of activity and inhibition of the hepatitis C virus RNA-dependent RNA polymerase. J. Biol. Chem. 285, 13685–13693.10.1074/jbc.M109.082206Search in Google Scholar

Rizo, J. and Südhof, T.C. (1998) C2-domains, structure and function of a universal Ca2+-binding domain. J. Biol. Chem. 273, 15879–15882.Search in Google Scholar

Rudd, J.J. and Franklin-Tong, V.E. (1999). Calcium signaling in plants. Cell. Mol. Life Sci. 55, 214–232.10.1007/s000180050286Search in Google Scholar

Schäffner, I., Rücknagel, K.-P., Mansfeld, J., and Ulbrich-Hofmann, R. (2002). Genomic structure, cloning and expression of two phospholipase D isoenzymes from white cabbage. Eur. J. Lipid Sci. Technol. 104, 79–87.10.1002/1438-9312(200202)104:2<79::AID-EJLT79>3.0.CO;2-CSearch in Google Scholar

Selvy, P.E., Lavieri, R.R., Lindsley, C.W., and Brown, H.A. (2011). Phospholipase D: enzymology, functionality, and chemical modulation. Chem. Rev. 111, 6064–6119.10.1021/cr200296tSearch in Google Scholar

Shannon, R.D. (1976). Revised effective ionic radii and systematic studies of interatomic distances in halides and chalcogenides. Acta Cryst. A 32, 751–767.10.1107/S0567739476001551Search in Google Scholar

Stumpe, S., König, S., and Ulbrich-Hofmann, R. (2007). Insights into the structure of plant α-type phopholipase D. FEBS J. 274, 2630–2640.10.1111/j.1742-4658.2007.05798.xSearch in Google Scholar

Tiwari, K. and Paliyath, G. (2011). Cloning, expression and functional characterization of the C2 domain from tomato phospholipase Dα. Plant Physiol. Biochem. 49, 18–32.10.1016/j.plaphy.2010.09.015Search in Google Scholar

Ulbrich-Hofmann, R., Lerchner, A., Oblozinsky, M., and Bezakova, L. (2005). Phospholipase D and its application in biocatalysis. Biotechnol. Lett. 27, 535–544.10.1007/s10529-005-3251-2Search in Google Scholar

Walters, J.D. and Johnson, J.D. (1990). Terbium as a luminescent probe of metal-binding sites in protein kinase C. J. Biol. Chem. 265, 4223–4226.10.1016/S0021-9258(19)39550-XSearch in Google Scholar

Zheng, L., Krishnamoorthi, R., Zolkiewski, M., and Wang, X. (2000). Distinct Ca2+ binding properties of novel C2 domains of plant phospholipase Dα and β. J. Biol. Chem. 275, 19700–19706.10.1074/jbc.M001945200Search in Google Scholar

Zheng, L., Shan, J., Krishnamoorthi, R., and Wang, X. (2002). Activation of plant phospholipase Dβ by phosphatidylinositol 4,5-bisphosphate: characterization of binding site and mode of action. Biochemistry 41, 4546–4553.10.1021/bi0158775Search in Google Scholar PubMed


Supplemental Material: The online version of this article (DOI 10.1515/hsz-2014-0112) offers supplementary material, available to authorized users.


Received: 2014-1-31
Accepted: 2014-3-13
Published Online: 2014-3-19
Published in Print: 2014-7-1

©2014 by Walter de Gruyter Berlin/Boston

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