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Ribosome-Inactivating Proteins from Abrus pulchellus

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Book cover Toxic Plant Proteins

Part of the book series: Plant Cell Monographs ((CELLMONO,volume 18))

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Abstract

Pulchellins are highly toxic type 2 ribosome-inactivating proteins (RIPs) expressed in the seeds of Abrus pulchellus tenuiflorus. Four pulchellin isoforms have been characterized, allowing their classification into two subgroups based on the toxicity levels and sugar-binding specificity. The residues involved in the catalytic mechanism are all conserved amongst the pulchellins, suggesting that the differential toxicity is related to variations in their B-chain behavior. In this chapter, insights into the successful production of active A- and B-chains via heterologous production in Escherichia coli and their assembly in vitro to produce an active heterodimer are discussed in some detail. Additionally, some features of subcellular sorting of native pulchellins are presented. Pulchellin comparative studies have contributed to the knowledge of the molecular bases of biochemical processes, such as sugar binding and toxicity, observed among RIPs.

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References

  • Bagaria A, Surendranath K, Ramagopal UA, Ramakumar S, Karande AA (2006) Structure–function analysis ans insights into the reduced toxicity os Abrus precatorius abrin. J Biol Chem 281:34465–34474

    Article  PubMed  CAS  Google Scholar 

  • Barbieri L, Ciani M, Girbes T, Liu WY, Van Damme EJ, Peumans WJ, Stirpe F (2004) Enzymatic activity of toxic and non-toxic type 2 ribosome-inactivating proteins. FEBS Lett 563:219–222

    Article  PubMed  CAS  Google Scholar 

  • Blakey DC, Thorpe PE (1988) Prevention of carbohydrate-mediated clearance of ricin-containing immunotoxins by the liver. Cancer Treat Res 37:457–473

    Article  PubMed  CAS  Google Scholar 

  • Castilho PV, Goto LS, Roberts LM, Araujo APU (2008) Isolation and characterization of four type 2 ribosome inactivating pulchellin isoforms from Abrus pulchellus seeds. FEBS J 275:948–959

    Article  PubMed  CAS  Google Scholar 

  • Chamberlain KL, Marshall RS, Jolliffe NA, Frigerio L, Ceriotti A, Lord JM, Roberts LM (2008) Ricin B chain targeted to the endoplasmic reticulum of tobacco protoplasts is degraded by a CDC48- and vacuole-independent mechanism. J Biol Chem 283:33276–33286

    Article  PubMed  CAS  Google Scholar 

  • Chambery A, Severino V, Stirpe F, Parente A (2007) Cloning and expression of the B chain of volkensin, type 2 ribosome inactivating protein from Adenia volkensii harms: co-folding with the A chain for heterodimer reconstitution. Protein Expr Purif 51:209–215

    Article  PubMed  CAS  Google Scholar 

  • Clark ED (2001) Protein refolding for industrial processes. Curr Opin Biotechnol 12:202–207

    Article  PubMed  CAS  Google Scholar 

  • D’Silva I, Vaidyanathan CS, Podder SK (1993) Ribosome-inactivating proteins and agglutinins from callus and suspension cultures of Ricinus communis L. and Abrus precatorius L. Plant Sci 94:161–172

    Article  Google Scholar 

  • Eck J, Langer M, Möckel B, Witthohn K, Zinke H, Lentzen H (1999) Characterization of recombinant and plant-derived mistletoe lectin and their B-chains. Eur J Biochem 265:788–797

    Article  PubMed  CAS  Google Scholar 

  • Endo Y, 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

    PubMed  CAS  Google Scholar 

  • Endo Y, Mitsui K, Motizuki M, 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 caused by the toxins. J Biol Chem 262:5908–5912

    PubMed  CAS  Google Scholar 

  • Frankel A, Roberts H, Gulick H, Afrin L, Vesely J, Willingham M (1994) Expression of ricin B chain in Spodoptera frugiperda. Biochem J 303:787–794

    PubMed  CAS  Google Scholar 

  • Frigerio L, Vitale A, Lord JM, Ceriotti A, Roberts LM (1998) Free ricin A chain, proricin, and native toxin have different cellular fates when expressed in tobacco protoplasts. J Biol Chem 273:14194–14199

    Article  PubMed  CAS  Google Scholar 

  • Girbés T, Ferreras JM, Arias FJ, Stirpe F (2004) Description, distribution, activity and phylogenetic relationship of ribosome-inactivating proteins in plant, fungi and bacteria. Mini Rev Med Chem 4:467–482

    Article  Google Scholar 

  • Goto LS, Beltramini LM, de Moraes DI, Moreira RA, de Araujo AP (2003) Abrus pulchellus type-2 RIP, pulchellin: heterologous expression and refolding of the sugar-binding B chain. Protein Expr Purif 31:12–18

    Article  PubMed  CAS  Google Scholar 

  • Goto LS, Castilho PV, Cominetti MR, Selistre-Araujo HS, Ulian Araujo AP (2007) Endocytosis of pulchellin and its recombinant B-chain into K-562 cells: binding and uptake studies. Biochim Biophys Acta 1770:1660–1666

    Article  PubMed  CAS  Google Scholar 

  • Gu Z, Su Z, Janson JC (2001) Urea gradient size-exclusion chromatography enhanced the yield of lysozyme refolding. J Chromatogr A 918:311–318

    Article  PubMed  CAS  Google Scholar 

  • Hartley MR, Lord JM (2004) Cytotoxic ribosome-inactivating lectins from plants. Biochim Biophys Acta 1701:1–14

    Article  PubMed  CAS  Google Scholar 

  • Hartley MR, Legname G, Osborn R, Chen Z, Lord MJ (1991) Single chain ribosome inactivating proteins from plants depurinate Escherichia coli 23S ribosomal RNA. FEBS Lett 290:65–68

    Article  PubMed  CAS  Google Scholar 

  • Hashimoto Y, Ono T, Goto M, Hatton TA (1998) Protein refolding by reversed micelles utilizing solid–liquid extraction technique. Biotechnol Bioeng 57:620–623

    Article  PubMed  CAS  Google Scholar 

  • Helmy M, Lombard S, Pieroni G (1999) Ricin RCA60: evidence of its phospholipase activity. Biochem Biophys Res Commun 258:252–255

    Article  PubMed  CAS  Google Scholar 

  • Hung CH, Lee MC, Chen JK, Lin JY (1994) Cloning and expression of three abrin A-chains and their mutants derived by site-specific mutagenesis in Escherichia coli. Eur J Biochem 219:83–87

    Article  PubMed  CAS  Google Scholar 

  • Jansen B, Vallera DA, Jaszcz WB, Nguyen D, Kersey JH (1992) Successful treatment of human acute T-cell leukemia in SCID mice using the anti-CD7-deglycosylated ricin A-chain immunotoxin DA7. Cancer Res 52:1314–1321

    PubMed  CAS  Google Scholar 

  • Jolliffe NA, Ceriotti A, Frigerio L, Roberts LM (2003) The position of the proricin vacuolar targeting signal is functionally important. Plant Mol Biol 5:631–641

    Article  Google Scholar 

  • Kadokura H, Katzen F, Beckwith J (2003) Protein disulfide bond formation in prokaryotes. Annu Rev Biochem 72:111–135

    Article  PubMed  CAS  Google Scholar 

  • Li XD, Liu WY, Niu CL (1996) Purification of a new ribosome-inactivating protein from the seeds of Cinnamomum porrectum and characterization of the RNA N-glycosidase activity of the toxic protein. Biol Chem 377:825–831

    PubMed  CAS  Google Scholar 

  • Lilie H, Schwarz E, Rudolph R (1998) Advances in refolding of proteins produced in E. coli. Curr Opin Biotechnol 9:497–501

    Article  PubMed  CAS  Google Scholar 

  • Maachupalli-Reddy J, Kelley BD, De Bernardez CE (1997) Effect of inclusion body contaminants on the oxidative renaturation of hen egg white lysozyme. Biotechnol Prog 13:144–150

    Article  PubMed  CAS  Google Scholar 

  • Magnusson S, Kjeken R, Berg T (1993) Characterization of two distinct pathways of endocytosis of ricin by rat liver endothelial cells. Exp Cell Res 205:118–125

    Article  PubMed  CAS  Google Scholar 

  • Navarro MVAS, Castilho PV, Araujo APU (Unpublished) Structural insights into the differential cytotoxicity presented by two isoforms of the RIP type 2 pulchellin

    Google Scholar 

  • Nielsen K, Boston RS (2001) Ribosome-inactivating proteins: a plant perspective. Annu Rev Plant Physiol Plant Mol Biol 52:785–816

    Article  PubMed  CAS  Google Scholar 

  • Olsnes S (2004) The history of ricin, abrin and related toxins. Toxicon 44:361–370

    Article  PubMed  CAS  Google Scholar 

  • Olsnes S, Pihl A (1973a) Different biological properties of the two constituent peptide chains of ricin, a toxic protein inhibiting protein synthesis. Biochemistry 12:3121–3126

    Article  PubMed  CAS  Google Scholar 

  • Olsnes S, Pihl A (1973b) Isolation and properties of abrin: a toxic protein inhibiting protein synthesis. Evidence for different biological functions of its two constituent-peptide chains. Eur J Biochem 35:179–185

    Article  PubMed  CAS  Google Scholar 

  • Parrotta JA (2001) Healing plants of peninsular India. CAB International, Wallingford

    Google Scholar 

  • Peumans WJ, van Damme EJ (1995) The role of lectins in plant defence. Histochem J 27:253–271

    Article  PubMed  CAS  Google Scholar 

  • Peumans WJ, Van Damme EJ, Barre A, Rouge P (2001) Classification of plant lectins in families of structurally and evolutionary related proteins. Adv Exp Med Biol 491:27–54

    Article  PubMed  CAS  Google Scholar 

  • Pevzner IB, Agapov II, Pfueller U, Pfueller K, Maluchenko NV, Moisenovich MM, Tonevitsky AG, Kirpichnikov MP (2005) Cloning and expression of mistletoe lectin III B-subunit. Biochemistry (Mosc) 70:306–315

    Article  CAS  Google Scholar 

  • Ramos MV, Mota DM, Teixeira CR, Cavada BS, Moreira RA (1998) Isolation and partial characterisation of highly toxic lectins from Abrus pulchellus seeds. Toxicon 36:477–484

    Article  PubMed  CAS  Google Scholar 

  • Ramos MV, Sampaio AH, Cavada BS, Calvete JJ, Grangeiro TB, Debray H (2001) Characterization of the sugar-binding specificity of the toxic lectins isolated from Abrus pulchellus seeds. Glycoconj J 18:391–400

    Article  PubMed  CAS  Google Scholar 

  • Rariy RV, Klibanov AM (1999) Protein refolding in predominantly organic media markedly enhanced by common salts. Biotechnol Bioeng 62:704–710

    Article  PubMed  CAS  Google Scholar 

  • Roberts LM, Smith DC (2004) Ricin: the endoplasmic reticulum connection. Toxicon 44:469–472

    Article  PubMed  CAS  Google Scholar 

  • Roberts LM, Tregear JW, Lord JM (1992) Molecular cloning of ricin. Targeted Diagn Ther 7:81–97

    PubMed  CAS  Google Scholar 

  • Rutenber E, Robertus JD (1991) Structure of ricin B-chain at 2.5 A resolution. Proteins 10:260–269

    Article  PubMed  CAS  Google Scholar 

  • Rutenber E, Ready M, Robertus JD (1987) Structure and evolution of ricin B chain. Nature 326:624–626

    Article  PubMed  CAS  Google Scholar 

  • Sehnke PC, Pedrosa L, Paul AL, Frankel AE, Ferl RJ (1994) Expression of active, processed ricin in transgenic tobacco. J Biol Chem 269:22473–22476

    PubMed  CAS  Google Scholar 

  • Shih RN, McDonald K, Jackman A, Girbés T, Iglesias R (1997) Bifunctional plant defense enzymes with chitinase and ribosome inactivating activities from Trichosanthes kirilowii cell cultures. Plant Sci 130:145–150

    Article  CAS  Google Scholar 

  • Silva ALC, Horta ACG, Moreira RA, Beltramini LM, Araujo APU (2003) Production of Abrus pulchellus ribosome-inactivating protein from seeds callus culture. Toxicon 41:841–849

    Article  PubMed  CAS  Google Scholar 

  • Silva AL, Goto LS, Dinarte AR, Hansen D, Moreira RA, Beltramini LM, Araujo AP (2005) Pulchellin, a highly toxic type 2 ribosome-inactivating protein from Abrus pulchellus. Cloning heterologous expression of A-chain and structural studies. FEBS J 272:1201–1210

    Article  PubMed  CAS  Google Scholar 

  • Simmons BM, Stahl PD, Russell JH (1986) Mannose receptor-mediated uptake of ricin toxin and ricin A chain by macrophages. Multiple intracellular pathways for a chain translocation. J Biol Chem 261:7912–7920

    PubMed  CAS  Google Scholar 

  • Sphyris N, Lord JM, Wales R, Roberts LM (1995) Mutational analysis of the Ricinus lectin B-chains. Galactose-binding ability of the 2 gamma subdomain of Ricinus communis agglutinin B-chain. J Biol Chem 270:20292–20297

    Article  PubMed  CAS  Google Scholar 

  • St John RJ, Carpenter JF, Randolph TW (1999) High pressure fosters protein refolding from aggregates at high concentrations. Proc Natl Acad Sci USA 96:13029–13033

    Article  PubMed  Google Scholar 

  • Stirpe F, Battelli MG (2006) Ribosome-inactivating proteins: progress and problems. Cell Mol Life Sci 63:1850–1866

    Article  PubMed  CAS  Google Scholar 

  • Stirpe F, Barbieri L, Battelli MG, Soria M, Lappi DA (1992) Ribosome-inactivating proteins from plants: present status and future prospects. Biotechnology 10:405–412

    Article  PubMed  CAS  Google Scholar 

  • Svinth M, Steighardt J, Hernandez R, Suh JK, Kelly C, Day P, Lord M, Girbes T, Robertus JD (1998) Differences in cytotoxicity of native and engineered RIPs can be used to assess their ability to reach the cytoplasm. Biochem Biophys Res Commun 249:637–642

    Article  PubMed  CAS  Google Scholar 

  • Tahirov TH, Lu TC, Liaw YC, Chen YL, Lin JY (1995) Crystal structure of abrin-a at 2.14 Å. J Mol Biol 250:354–367

    Article  PubMed  CAS  Google Scholar 

  • Tonevitsky A, Toptygin A, Agapov I, Pfueller U, Frankel A (1994) Renaturated ricin toxin B chain made in Escherichia coli is soluble, stable, and biologically active. Biochem Mol Biol Int 32:1139–1146

    PubMed  CAS  Google Scholar 

  • Tsumoto K, Ejima D, Kumagai I, Arakawa T (2003) Practical considerations in refolding proteins from inclusion bodies. Protein Expr Purif 28:1–8

    Article  PubMed  CAS  Google Scholar 

  • VanDamme EJM, Peumans WJ, Barre A, Rougé P (1998) Plant lectins: a composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles. Crit Rev Plant Sci 17:575–692

    Article  CAS  Google Scholar 

  • Wales R, Richardson PT, Roberts LM, Woodland HR, Lord JM (1991) Mutational analysis of the galactose binding ability of recombinant ricin B chain. J Biol Chem 266:19172–19179

    PubMed  CAS  Google Scholar 

  • Wang P, Tumer NE (2000) Virus resistance mediated by ribosome inactivating proteins. Adv Virus Res 55:325–355

    Article  PubMed  CAS  Google Scholar 

  • Wood KA, Lord JM, Wawrzynczak EJ, Piatak M (1991) Preproabrin: genomic cloning, characterization and the expression of the A-chain in Escherichia coli. Eur J Biochem 198:723–732

    Article  PubMed  CAS  Google Scholar 

  • Zardeneta G, Horowitz PM (1994) Detergent, liposome, and micelle-assisted protein refolding. Anal Biochem 223:1–6

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Instituto de Física de São Carlos, Universidade de São Paulo, 369, 13560-970, São Carlos, SP, Brazil

    Ana Paula Ulian Araújo, Priscila Vasques Castilho & Leandro Seiji Goto

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  1. Ana Paula Ulian Araújo
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  2. Priscila Vasques Castilho
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  3. Leandro Seiji Goto
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Correspondence to Ana Paula Ulian Araújo .

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Editors and Affiliations

  1. Dept. Biological Sciences, University of Warwick, Coventry, CV47AL, United Kingdom

    J. Michael Lord

  2. Dept. Biological Sciences, University of Warwick, Coventry, CV47AL, United Kingdom

    Martin R. Hartley

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Araújo, A.P.U., Castilho, P.V., Goto, L.S. (2010). Ribosome-Inactivating Proteins from Abrus pulchellus . In: Lord, J., Hartley, M. (eds) Toxic Plant Proteins. Plant Cell Monographs, vol 18. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-12176-0_7

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