Production of Ribosome-inactivating Protein from Hairy Root Cultures of Luffa Cylindrica (L.) Roem

Overview

Journal Plant Cell Rep

Publisher Springer

Date 2013 Nov 2

PMID 24178273

Citations 1

Authors

L S di Toppi

P Gorini

G Properzi

L Barbieri

L Spano

Affiliations

Soon will be listed here.

Abstract

Transformed root lines of Luffa cylindrica (L.) Roem. (Cucurbitaceae) were established by inoculation of in vitro grown plantlets with wild type Agrobacterium rhizogenes strain 1855. Cloned lines of hairy roots were tested for the presence of ribosome-inactivating proteins; crude extracts inhibited protein synthesis in a reaction mixture based on rabbit reticulocyte lysate. Inhibitory activity increased during culture period, reaching a maximum value in the stationary phase. No activity could be detected in the culture medium, nor in extracts from callus and/or suspension cultures. A ribosome-inactivating protein having specific activity of 62,100 U mg protein(-1) and a molecular mass of 26-28,000 Da was purified to homogeneity. The protein showed N-glycosidase activity on rat liver ribosomes. The results demonstrate that hairy root cultures can be successfully utilized for the in vitro production of ribosome-inactivating proteins.

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References

Barbieri L, Ferreras J, Barraco A, Ricci P, Stirpe F . Some ribosome-inactivating proteins depurinate ribosomal RNA at multiple sites. Biochem J. 1992; 286 ( Pt 1):1-4. PMC: 1133008. DOI: 10.1042/bj2860001. View

Gamborg O, Miller R, Ojima K . Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res. 1968; 50(1):151-8. DOI: 10.1016/0014-4827(68)90403-5. View

Ng T, Wong R, Yeung H . Two proteins with ribosome-inactivating, cytotoxic and abortifacient activities from seeds of Luffa cylindrica roem (Cucurbitaceae). Biochem Int. 1992; 27(2):197-207. View

Misawa M, Hayashi M, Tanaka H . Production of plant virus inhibitor by Phytolacca americana suspension culture. Biotechnol Bioeng. 1975; 17(9):1335-47. DOI: 10.1002/bit.260170908. View

Islam M, Kung S, Kimura Y, Funatsu G . N-acetyl-D-glucosamine-asparagine structure in ribosome-inactivating proteins from the seeds of Luffa cylindrica and Phytolacca americana. Agric Biol Chem. 1991; 55(5):1375-81. View

Hamill J, Prescott A, Martin C . Assessment of the efficiency of cotransformation of the T-DNA of disarmed binary vectors derived from Agrobacterium tumefaciens and the T-DNA of A. rhizogenes. Plant Mol Biol. 2013; 9(6):573-84. DOI: 10.1007/BF00020534. View

Ikeda T, Takanami Y, Imaizumi S, Matsumoto T, Mikami Y, Kubo S . Formation of anti-plant viral protein by Mirabilis jalapa L. cells in suspension culture. Plant Cell Rep. 2013; 6(3):216-8. DOI: 10.1007/BF00268483. View

Islam M, Hirayama H, Funatsu G . Complete amino acid sequence of luffin-b, a ribosome-inactivating protein from sponge gourd (Luffa cylindrica) seeds. Agric Biol Chem. 1991; 55(1):229-38. View

Savary B, Flores H . Biosynthesis of defense-related proteins in transformed root cultures of Trichosanthes kirilowii Maxim. var japonicum (Kitam.). Plant Physiol. 1994; 106(3):1195-204. PMC: 159649. DOI: 10.1104/pp.106.3.1195. View

10.

Barbieri L, Battelli M, Stirpe F . Ribosome-inactivating proteins from plants. Biochim Biophys Acta. 1993; 1154(3-4):237-82. DOI: 10.1016/0304-4157(93)90002-6. View

11.

Barbieri L, Bolognesi A, Cenini P, Falasca A, Minghetti A, Garofano L . Ribosome-inactivating proteins from plant cells in culture. Biochem J. 1989; 257(3):801-7. PMC: 1135659. DOI: 10.1042/bj2570801. View

12.

Bonness M, Mabry T . Tissue culture of endod (Phytolacca dodecandra L'Herit): growth and production of ribosome-inactivating proteins. Plant Cell Rep. 2013; 11(2):66-70. DOI: 10.1007/BF00235255. View

13.

Inomata S, Yokoyama M, Gozu Y, Shimizu T, Yanagi M . Growth pattern and ginsenoside production of Agrobacterium-transformed Panax ginseng roots. Plant Cell Rep. 2013; 12(12):681-6. DOI: 10.1007/BF00233419. View

14.

Islam M, Nishida H, Funatsu G . Complete amino acid sequence of luffin-a, a ribosome-inactivating protein from the seeds of Luffa cylindrica. Agric Biol Chem. 1990; 54(5):1343-5. View

15.

Thomsen S, Hansen H, Nyman U . Ribosome-inhibiting proteins from in vitro cultures of Phytolacca dodecandra. Planta Med. 1991; 57(3):232-6. DOI: 10.1055/s-2006-960080. View

16.

Kalb Jr V, BERNLOHR R . A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977; 82(2):362-71. DOI: 10.1016/0003-2697(77)90173-7. View

17.

Watanabe K, Suemasu Y, Funatsu G . Identification of lysine residue at or near active site of luffin-a, a ribosome-inactivating protein from seeds of Luffa cylindrica. J Biochem. 1989; 106(6):977-81. DOI: 10.1093/oxfordjournals.jbchem.a122985. View

18.

Spano L, Pomponi M, Costantino P, van Slogteren G, Tempe J . Identification of T-DNA in the root-inducing plasmid of the agropine type Agrobacterium rhizogenes 1855. Plant Mol Biol. 2013; 1(4):291-300. DOI: 10.1007/BF00027560. View

19.

Ferreras J, Barbieri L, Girbes T, Battelli M, Rojo M, Arias F . Distribution and properties of major ribosome-inactivating proteins (28 S rRNA N-glycosidases) of the plant Saponaria officinalis L. (Caryophyllaceae). Biochim Biophys Acta. 1993; 1216(1):31-42. DOI: 10.1016/0167-4781(93)90034-b. View

20.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View