Certain protein transducing agents convert translocated proteins into cell killers

• Authors: Siergiej Tcherniuk , Anne-Laure Fiser , Madiha Derouazi , Bertrand Toussaint , Yan Wang , Izabela Wojtal , Eisaku Kondo , Ewa Szolajska , Jadwiga Chroboczek
• Languages of publication: EN

Abstracts

EN

The majority of proteins are unable to translocate into the cell interior. Hence for peptide- and protein-based therapeutics a direct intracytoplasmic delivery with the aid of transducing agents is an attractive approach. We wanted to deliver to the cell interior a putatively cytotoxic protein VPg. Protein transduction was achieved in vitro with three different commercial products. However, in our hands, delivery of various control proteins without known deleterious effects, as well as of protein VPg, always induced cell death. Finally, we used a novel transducing peptide Wr-T, which was not toxic to cultured cells, even in a quite large range of concentrations. Most importantly, control protein delivered to cells in culture did not display any toxicity while VPg protein exerted a strong cytotoxic effect. These data show that results obtained with cell-penetrating agents should be interpreted with caution.

Keywords

EN

cytotoxic protein; cell transduction; protein delivery; cell translocation; transducing peptides

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2012
• Volume: 59
• Issue: 3
• Pages: 433-439

Dates

published

2012

received

2012-07-03

revised

2012-08-23

accepted

2012-09-02

(unknown)

2012-09-03

Contributors

author

Siergiej Tcherniuk

• Institute of Structural Biology, CNRS-CEA-UJF, UMR 5075, Grenoble, France

author

Anne-Laure Fiser

• Institute of Structural Biology, CNRS-CEA-UJF, UMR 5075, Grenoble, France

author

Madiha Derouazi

• TheREx, TIMC-IMAG, UMR5525 CNRS/Université Joseph Fourier Faculté de Médecine, Bat. J Roget, La Tronche, France

author

Bertrand Toussaint

• TheREx, TIMC-IMAG, UMR5525 CNRS/Université Joseph Fourier Faculté de Médecine, Bat. J Roget, La Tronche, France

author

Yan Wang

• TheREx, TIMC-IMAG, UMR5525 CNRS/Université Joseph Fourier Faculté de Médecine, Bat. J Roget, La Tronche, France

author

Izabela Wojtal

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

author

Eisaku Kondo

• Department of Pathology, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan

author

Ewa Szolajska

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

author

Jadwiga Chroboczek

• Institute of Structural Biology, CNRS-CEA-UJF, UMR 5075, Grenoble, France

References

• Baltanás FC, Weruaga E, Valero J, Recio JS, Alonso JR (2008) Albumin attenuates DNA damage in primary-cultured neurons. Neurosci Lett 450: 23-26.
• Bildstein L, Dubernet C, Couvreur P (2011) Prodrug-based intracellular delivery of anticancer agents. Adv Drug Deliv Rev 63: 3-23.
• Cassinelli G, Lanzi C, Petrangolini G, Tortoreto M, Pratesi G, Cuccuru G et al. (2006) Inhibition of c-Met and prevention of spontaneous metastatic spreading by the 2-indolinone RPI-1. Mol Cancer Ther 5: 2388-2397.
• Cortes C, Rzomp KA, Tvinnereim A, Scidmore MA, Wizel B (2007) Chlamydia pneumoniae inclusion membrane protein Cpn0585 interacts with multiple Rab GTPases. Infect Immun 75: 5586-5596.
• Chowdhury D, Lieberman J. (2008) Death by a thousand cuts: granzyme pathways of programmed cell death. Annu Rev Immunol 26: 389-420.
• Cormack BP, Valdivia RH, Falkow S. (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173: 33-38
• Cortes C, Rzomp KA, Tvinnereim A, Scidmore MA, Wizel B (2007) Chlamydia pneumoniae inclusion membrane protein Cpn0585 interacts with multiple Rab GTPases. Infect Immun 75: 5586-5596.
• Ceckova M, Vackova Z, Radilova H, Libra A, Buncek M, Staud F (2008) Effect of ABCG2 on cytotoxicity of platinum drugs: interference of EGFP. Toxicol In vitro 22: 1846-1852.
• Dalkara D, Zuber G, Behr JP (2004) Intracytoplasmic delivery of anionic proteins. Mol Ther 9: 964-969.
• Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19: 5720-578.
• Dietz GP (2010) Cell-penetrating peptide technology to deliver chaperones and associated factors in diseases and basic research. Curr Pharm Biotechnol 11: 167-174.
• Kondo E, Seto M, Yoshikawa K, Yoshino T (2004) Highly efficient delivery of p16 antitumor peptide into aggressive leukemia/lymphoma cells using a novel transporter system. Mol Cancer Ther 3: 1623-1630.
• Kondo E, Tanaka T, Miyake T, Ichikawa T, Hirai M, Adachi M et al. (2008) Potent synergy of dual antitumor peptides for growth suppression of human glioblastoma cell lines. Mol Cancer Ther 7: 1461-1471.
• Kroemer G, Levine B (2008) Autophagic cell death: the story of a misnomer. Nat Rev Mol Cell Biol 9: 1004-1010.
• Kurono M, Fujii A, Murata M, Fujitani B, Negoro T (2006) Stereospecific recognition of a spirosuccinimide type aldose reductase inhibitor (AS-3201) by plasma proteins: a significant role of specific binding by serum albumin in the improved potency and stability. Biochem Pharmacol 71: 338-353.
• Lucast LJ, Batey RT, Doudna JA (2001) Large-scale purification of a stable form of recombinant tobacco etch virus protease. Biotechniques 30: 544-546.
• Lundberg M, Wikström S, Johansson M (2003) Cell surface adherence and endocytosis of protein transduction domains. Mol Ther 8: 143-150.
• Liu HS, Jan MS, Chou CK, Chen PH, Ke NJ (1999) Is green fluorescent protein toxic to the living cells? Biochem Biophys Res Commun 260: 712-717.
• Lu H, Hou Q, Zhao T, Zhang H, Zhang Q, Wu L, Fan Z (2006) Granzyme M directly cleaves inhibitor of caspase-activated DNase (CAD) to unleash CAD leading to DNA fragmentation. J Immunol 177: 1171-1178.
• Mo RH, Zaro JL, Ou JH, Shen WC (2012) Effects of Lipofectamine 2000/siRNA complexes on autophagy in hepatoma cells. Mol Biotechnol 51: 1-8.
• Meredith SC (2005) Protein denaturation and aggregation: Cellular responses to denatured and aggregated proteins. Ann N Y Acad Sci 1066: 181-221.
• Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.
• Reales E, Mora-López F, Rivas V, García-Poley A, Brieva JA, Campos-Caro A (2005) Identification of soluble N-ethylmaleimide-sensitive factor attachment protein receptor exocytotic machinery in human plasma cells: SNAP-23 is essential for antibody secretion. J Immunol 175: 6686-6693.
• Salomons FA, Menéndez-Benito V, Böttcher C, McCray BA, Taylor JP, Dantuma NP (2009) Selective accumulation of aggregation-prone proteasome substrates in response to proteotoxic stress. Mol Cell Biol 29: 1774-1785.
• Scarlatti F, Granata R, Meijer AJ, Codogno P (2009) Does autophagy have a license to kill mammalian cells? Cell Death Differ 16: 12-20.
• Schmalz G, Hoffmann M, Weis K, Schweikl H (2000) Influence of albumin and collagen on the cell mortality evoked by zinc oxide-eugenol in vitro. J Endod 26: 284-287.
• Shi J, Karlsson HL, Johansson K, Gogvadze V, Xiao L, Li J, Burks T, Garcia-Bennett A, Uheida A, Muhammed M, Mathur S, Morgenstern R, Kagan VE, Fadeel B (2012) Microsomal glutathione transferase 1 protects against toxicity induced by silica nanoparticles but not by zinc oxide nanoparticles. ACS Nano 6: 1925-1938.
• Valkonen M, Ward M, Wang H, Penttilä M, Saloheimo M (2003) Improvement of foreign-protein production in Aspergillus niger var. awamori by constitutive induction of the unfolded-protein response. Appl Environ Microbiol 69: 6979-6986.
• Weill CO, Biri S, Erbacher P (2008) Cationic lipid-mediated intracellular delivery of antibodies into live cells. Biotechniques 44: Pvii-Pxi.
• Zhang G, Gurtu V, Kain SR (1996) An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells. Biochem Biophys Res Comm 227: 707-711.
• Zhao T, Zhang H, Guo Y, Zhang Q, Hua G, Lu H et al. (2007) Granzyme K cleaves the nucleosome assembly protein SET to induce single-stranded DNA nicks of target cells. Cell Death Differ 14: 489-499.