Home An apoptotic inducer, aralin, is a novel type II ribosome-inactivating protein from Aralia elata
Article
Licensed
Unlicensed Requires Authentication

An apoptotic inducer, aralin, is a novel type II ribosome-inactivating protein from Aralia elata

  • Makoto Tomatsu , Toshiya Kondo , Takafumi Yoshikawa , Takashi Komeno , Naomi Adachi , Yasushi Kawasaki , Akira Ikuta and Fumio Tashiro
Published/Copyright: June 1, 2005
Biological Chemistry
From the journal Volume 385 Issue 9

Abstract

We recently found that aralin, a novel cytotoxic protein consisting of two subunits, from Aralia elata selectively induces apoptosis in transformed cells as compared to normal cells. Here we report that aralin is a lectin specific for galactose (Gal) and its derivatives, and possesses RNA N-glycosidase activity as a new type II ribosome-inactivating protein (RIP). The RNA N-glycosidase activity of aralin was detected in cell-free and whole cell systems by the generation of an R-fragment from 28S rRNA. Coinciding with appearance of the R-fragment in aralin-treated cells, significant inhibition of protein synthesis was observed prior to the onset of apoptosis. Aralin-evoked cell death was efficiently repressed by the addition of Gal and its derivatives. Interestingly, melibiose preferentially protected normal cells from apoptosis as compared with transformed cells. Using rhodamine-coupled aralin, the aralin receptor could be clearly detected around the cell surface of transformed cells, but to a lesser extent on normal cells. Receptor binding was suppressed by Gal. These results indicate that aralin is incorporated into cells via its Gal-containing cell surface receptor and induces apoptosis through its RIP activity. Moreover, the expression level and/or structural changes of the aralin receptor may affect the sensitivity toward aralin.

:

References

Barbieri, L., Battelli, M.G., and Stirpe, F. (1993). Ribosome-inactivating proteins from plants. Biochim. Biophys. Acta1154, 237–282.10.1016/0304-4157(93)90002-6Search in Google Scholar

Chomczynski, P., and Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem.162, 156–159.10.1016/0003-2697(87)90021-2Search in Google Scholar

Cummings, R.D., and Kornfeld, S. (1982). Fractionation of asparagine-linked oligosaccharides by serial lectin-agarose affinity chromatography. J. Biol. Chem.257, 11235–11240.10.1016/S0021-9258(18)33747-5Search in Google Scholar

Endo, Y., and Tsurugi, K. (1987). RNA N-glycosidase activity of ricin A-chain: mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. J. Biol. Chem.262, 8128–8130.10.1016/S0021-9258(18)47538-2Search in Google Scholar

Endo, Y., Mitsui, K., Motizuki, M., and Tsurugi, K. (1987). The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes: the site and the characteristics of the modification in 28S ribosomal RNA caused by the toxins. J. Biol. Chem.262, 5908–5912.10.1016/S0021-9258(18)45660-8Search in Google Scholar

Girbés, T., Ferreras, J.M., Iglesias, R., Citores, L., de Torre, C., Carbajales, M.L., Jiménez, P., de Benito, F.M., and Muñoz, R. (1996). Recent advances in the uses and applications of ribosome-inactivating proteins from plants. Cell. Mol. Biol.42, 461–471.Search in Google Scholar

Girbés, T., Ferreras, J.M., Arias, F.J., Muñoz, R., Iglesias, R., Jimenez, P., Rojo, M.A., Arias, Y., Perez, Y., Benitez, J., Sanchez, D., and Gayoso, M.J. (2003). Non-toxic type 2 ribosome-inactivating proteins (RIPs) from Sambucus: occurrence, cellular and molecular activities and potential uses. Cell. Mol. Biol.49, 537–545.Search in Google Scholar

Griffiths, G.D., Leek, M.D., and Gee, D.J. (1987). The toxic plant proteins ricin and abrin induce apoptotic change in mammalian lymphoid tissues and intestine. J. Pathol.151, 221–229.10.1002/path.1711510310Search in Google Scholar

Hardy, M.R., Townsend, R.R., and Lee, Y.C. (1988). Monosaccharide analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection. Anal. Biochem.170, 54–62.10.1016/0003-2697(88)90089-9Search in Google Scholar

Iida, N., Sugiyama, A., Myoubudani, H., Inoue, K., Sugamata, M., Ihara, T., Ueno, Y., and Tashiro, F. (1998). Suppression of arachidonic acid cascade-mediated apoptosis in aflatoxin B1–induced rat hepatoma cells by glucocorticoids. Carcinogenesis19, 1191–1202.10.1093/carcin/19.7.1191Search in Google Scholar PubMed

Kawabata, Y., Tashiro, F., and Ueno, Y. (1982). Synthesis of a specific protein induced by zearalenone and its derivatives in rat uterus. J. Biochem.91, 801–808.10.1093/oxfordjournals.jbchem.a133767Search in Google Scholar PubMed

Kondo, T., Yoshikawa, T., Ogihara, Y., and Mizukami, H. (2002). Fusion with maltose-binding protein (MBP) affects neither RNA N-glycosidase activity nor immunogenicity of karasurin- A, a ribosome-inactivating protein from Trichosanthes kirilowii var. japonica. Biotechnol. Lett.24, 1117–1124.10.1023/A:1016035205596Search in Google Scholar

Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227, 680–685.10.1038/227680a0Search in Google Scholar

Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem.193, 265–275.10.1016/S0021-9258(19)52451-6Search in Google Scholar

Magnússon, S., and Berg, T. (1993). Endocytosis of ricin by rat liver cells in vivo and in vitro is mainly mediated by mannose receptors on sinusoidal endothelial cells. Biochem. J.291, 749–755.10.1042/bj2910749Search in Google Scholar

Martin, S.J., Lennon, S.V., Bonham, A.M., and Cotter, T.G. (1990). Induction of apoptosis (programmed cell death) in human leukemic HL-60 cells by inhibition of RNA or protein synthesis. J. Immunol.145, 1859–1867.10.4049/jimmunol.145.6.1859Search in Google Scholar

Nielsen, K., and Boston, R.S. (2001). Ribosome-inactivating proteins: a plant perspective. Annu. Rev. Plant Physiol. Plant Mol. Biol.52, 785–816.10.1146/annurev.arplant.52.1.785Search in Google Scholar

Olsnes, S. (1978). Ricin and ricinus agglutinin, toxic lectins from castor bean. Methods Enzymol.50, 330–335.10.1016/0076-6879(78)50037-2Search in Google Scholar

Olsnes, S., Saltvedt, E., and Pihl, A. (1974). Isolation and comparison of galactose-binding lectins from Abrus precatorius and Ricinus communis. J. Biol. Chem.249, 803–810.10.1016/S0021-9258(19)43000-7Search in Google Scholar

Peumans, W.J., Hao, Q., and van Damme, E.J.M. (2001). Ribosome-inactivating proteins from plants: more than RNA N-glycosidases? FASEB J.15, 1493–1506.10.1096/fj.00-0751revSearch in Google Scholar

Sandvig, K., Olsnes, S., Brown, J.E., Petersen, O.W., and van Deurs, B. (1989). Endocytosis from coated pits of Shiga toxin: a glycolipid-binding protein from Shigella dysenteriae 1. J. Cell Biol.108, 1331–1343.10.1083/jcb.108.4.1331Search in Google Scholar

Sandvig, K., and van Deurs, B. (2000). Entry of ricin and Shiga toxin into cells: molecular mechanisms and medical perspective. EMBO J.19, 5943–5950.10.1093/emboj/19.22.5943Search in Google Scholar

Skilleter, D.N., Price, R.J., and Thorpe, P.E. (1985). Modification of the carbohydrate in ricin with metaperiodate and cyanoborohydride mixtures: effect on binding, uptake and toxicity to parenchymal and non-parenchymal cells of rat liver. Biochim. Biophys. Acta842, 12–21.10.1016/0304-4165(85)90287-9Search in Google Scholar

Staehelin, T., and Falvey, A.K. (1971). Isolation of mammalian ribosomal subunits active in polypeptide synthesis. Methods Enzymol.20, 433–446.10.1016/S0076-6879(71)20047-1Search in Google Scholar

Tamura, T., Sadakata, N., Oda, T., and Muramatsu, T. (2002). Role of zinc ions in ricin-induced apoptosis in U937 cells. Toxicol. Lett.132, 141–151.10.1016/S0378-4274(02)00078-4Search in Google Scholar

Tazzari, P.L., Polito, L., Bolognesi, A., Pistillo, M.P., Capanni, P., Palmisano, G.L., Lemoli, R.M., Curti, A., Biancone, L., Camussi, G. et al. (2001). Immunotoxins containing recombinant anti-CTLA-4 single-chain fragment variable antibodies and saporin: in vitro results and in vivo effects in an acute rejection model. J. Immunol.167, 4222–4229.10.4049/jimmunol.167.8.4222Search in Google Scholar

Thorpe, P.E., Detre, S.I., Foxwell, B.M., Brown, A.N., Skilleter, D.N., Wilson, G., Forrester, J.A., and Stirpe, F. (1985). Modification of the carbohydrate in ricin with metaperiodate-cyanoborohydride mixtures: effects on toxicity and in vivo distribution. Eur. J. Biochem.147, 197–206.10.1111/j.1432-1033.1985.tb08737.xSearch in Google Scholar

Tomatsu, M., Kameyama, M.O., and Shibamoto, N. (2003). Aralin, a new cytotoxic protein from Aralia elata, induces apoptosis in human cancer cells. Cancer Lett.199, 19–25.10.1016/S0304-3835(03)00348-3Search in Google Scholar

Tomatsu, M., Mujin, T., Shibamoto, N., Tashiro, F., and Ikuta, A. (2004). Production of aralin, a selective cytotoxic lectin against human transformed cells, in callus culture of Aralia elata. Planta Med.70, 469–471.Search in Google Scholar

Toyokawa, S., Takeda, T., and Ogihara, Y. (1991). Isolation and characterization of a new abortifacient protein, karasurin, from root tubers of Trichosanthes kirilowii Max. var. japonicam Kitam. Chem. Pharm. Bull.39, 716–719.10.1248/cpb.39.716Search in Google Scholar PubMed

van Oijen, M.G., and Preijers, F.W. (1997). Rationale for the use of immunotoxins in the treatment of HIV-infected humans. J. Drug Target.5, 75–91.Search in Google Scholar

Willey, R.G. and Oeltmann, T.N. (1991). Ricin and related plant toxins: Mechanism of action and neurobiological applications. In: Handbook of Natural Toxins, volume 6, R.F. Keeler and A.T. Tu, eds. (New York, USA: Marcel Dekker Inc.), pp. 243–268.Search in Google Scholar

Published Online: 2005-06-01
Published in Print: 2004-09-01

© Walter de Gruyter

Downloaded on 17.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/BC.2004.107/html
Scroll to top button