Puji Astuti, Sudjadi ., Sismindari .


Ribosome Inactivating Proteins (RIPs) are compounds isolated from plants with ability to inhibit protein synthesis. The inhibition of protein synthesis is due to inactivation of ribosomal RNA through a site-specific  deadenylation  mediated  by  RNA  N-glycosidase. Reportedly, RIPs mainly possess wide range of bioactivity including antiviral  activity  against  plant  infections.  Other  activities  of  RIP were  as  abortifacien,  antivirus  and  anticancer.  This  study  was aimed  to  isolate  and  characterize  the  optimum  conditions  for inducing  the  expression  of  recombinant  RIPs  isolated  from  the leaves  of  Mirabilis  Jalapa  L.  We  have  been  successfully  isolated several  RIPs  and  engineered  these  proteins  to  be  expressed  in  E. coli. These recombinant proteins were obtained by screening cDNA library  originated  from  the  mRNA  of  Mirabilis  jalapa  L  leaves,  and inserted  into  pUC19  carrying  lacZ  gene.  The  presence  of recombinant  plasmid  was  tested  by  using  α-complementation assay. Many RIPs have been isolated from plants and these proteins express  enzymatic  activity  by  cutting  supercoiled  double  stranded DNA. One RIP namely rMJC15310 was obtained from this study and the  proteins  having  ~  8kb  in  size,  cut  the  supercoiled  DNA  into linear  form  at  the  concentration  as  low  as  5  µg.  The  ability  to  cut supercoiled  DNA  increased  on  inducing  its  expression  with  0.4% IPTG.

Key words:   Ribosome  Inactivating  Proteins  (RIP),  IPTG,  Mirabilis  jalapa L., recombinant protein


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Barbieri, L., Batteli, M.A., Stirpe, F., 1993, Ribosome inactivating proteins from Plants, Biochem et Biophys. Acta, 405-410

Barbieri, L., Valbonesi, P., Bonora, E., Gorini, P., Bolognesi, A., and Stirpe, F., 1997, Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins : effect on DNA, RNA and poly(A). Nucleic Acids Res. 25, 518–522

Barberi, L., Polito, L., Bolognesi, A., Ciani, M., Pelosi, E., Farini, V., Jha, A.K., Sharma,N., Vivano, J.M., Chambery, A., Parente, A., and Stirpe, F., 2006,

Ribosomeinactiva-ting proteins in edible plants and purification and characterization of a new ribosomeinactivating protein from Cucurbita moschata, Biochim. Biophys. Acta, 170: 783-792.

Batteli, M.G., 2004, Cytotoxicity and toxicity to animals and humans of ribosomeinactivating proteins. Mini Rev Med Chem, 4(5):513-21.

Endo, Y., and Tsurugi, K.,1987, RNA Nglycosidase activity of ricin A-chain. Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. J. Biol. Chem. 262, 8128–8130

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

Li, M.X., Yeung, H.-W., Pan, L.P. and Chan, S.I.,1991, Nucleic Acids Res. 19, 63096312.

Lin, J. Y., Tserng, K. Y., Chen, C. C., Lin, L. T., and Tung, T. C., 1970, Abrin and ricin: new anti-tumour substances. Nature (London) 227, 292–293

Ling, J., Lui, W., and Wang, T.P., 1994, Cleavage of supercoiled double stranded DNA by several ribosome inactivating proteins in vitro, FEBS Letters, 345: 143-146.

Lodish, H., Berk, A., Zipursky, S.L., Matsudaira, P., Baltimore, D., Darnell., J.E., 2000, Molecular Cell Biology, 4th Ed., 244, W.H. Freeman and Company, New York.

Narayanan, S., K. Surendranath, N. Bora, A. Surolia, and A.A. Karande. 2005. Ribosome inactivating proteins and apoptosis. FEBS Lett. 579:1324-1331.

Peumans,W., Hoa,Q., and van Damme,E., 2001, Ribosome-inactivating proteins from plants: more than RNA Nglycosidase? J FASEB, 15: 1493-1506

Qu,X. and Qing,L., 2004, Abrin induces cell apoptosis by cytochrom c release and caspaseactivation,J.Bio.Mic.37(4),445-453.

Roulean, C., Curiel, M., Weber, W., Smale, R., Kurtzberg, L., Masearello, J., Berger, C., Waller, G., Bagley, R., Honma, N., Hasegawa,K., Ishida,I., Kataoka,S., Mehraenin,K., Horten, Miller,G., and Teicher,B.H., 2008, Endosialin protein expression and therapeutic target potential in human solid tumors: Sarcoma versus carcinoma, Clin. Can.Res.14, 7223.

Sambrook,J., Fritsch,E.F., Maniatis,T., 1989, Molecular Cloning : a Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, New York.

Sha, O., Yew, D.T., Ng, T.B., Yuan, L., Kwong, W.H., 2010, Different in vitro toxicities of structurally similar type I ribosomeinactivating proteins (RIPs). Toxicol In Vitro. 24 (4):1176-82

Sismindari, Husaana,A., Mubarika,S., 1998, In vitro cleavage of Supercoiled DNA by Annona squamosa extract, Indonesian J. Pharm., 9 (4), 146-152.

Sismindari, Mubarika S, and Sudjadi, 2001, Selective cytotoxic effects on cancer celllines of protein fraction isolated from Annona squamosa and containing ribosome-inactivating proteins, J.Biotech. 459 – 462.

Sismindari, Handayani A., Yulia S., and Chandra E., 2002, Potent effects of protein extract isolated from Erythrina fusca Lour leaves on cancer cell-lines,. J.Biotech. 559-564

Stirpe, F., Barbieri, R., Battteli, MG., Soria, M., and Iappi, D.A., 1992, RIPs from Plants: Present status and Future prospect, Rev.Biotech 10: 105-109

Stirpe,F., 2004, Ribosome-inactivating proteins, Toxicon, 44, 371 – 383

Sudjadi, Sismindari, Astuti,P., 2010, Kloning gena RIP penyandi Ribosome Inactivating protein dari daun Mirabilis jalapa L dengan pendekatan cDNA library, Proceeding Kongres Ilmiah IAI XVIII dan Rapat Kerja Nasional 2010, Makasar.

Wang, P., Zoubenko, O., and Tumer, N. E.,1998, Reduced toxicity and broad spectrum resistance to viral and fungal infection in transgenic plants expressing pokeweed antiviral protein II. Plant Mol. Biol. 38, 957–964



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