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Research article

Biotechnology and Bioengineering (BB)  |  Enzyme, Protein Engineering, and Metabolic Engineering

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand

Bich Hang Do 1, Minh Tan Nguyen 1, Jung-A Song 1, Sangsu Park 1, Jiwon Yoo 1, Jaepyeong Jang 1, Sunju Lee 1, Seoungjun So 1, Yejin Yoon 1, Inki Kim 2, Kyungjin Lee 2, Yeon Jin Jang and Han Choe 1*

1Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, Republic of Korea, 2Department of Convergence Medicine, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, Republic of Korea, 3Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea

Received: May 25, 2017; Accepted: September 25, 2017

J. Microbiol. Biotechnol. 2017; 27(12): 2156-2164

Published December 28, 2017 https://doi.org/10.4014/jmb.1705.05070

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b’a’ domain of protein disulfide isomerase (PDIb’a’), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was 0.4 EU/μg, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an EC50 and Hill coefficient of 0.6 ± 0.03 nM and 2.41 ± 0.15, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

Keywords

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), expression, purification, E. coli, cytotoxic activity, HeLa cells

References

  1. Wang S, El-Deiry WS. 2003. TRAIL and apoptosis induction by TNF-family death receptors. Oncogene 22: 8628-8633.
    Pubmed CrossRef
  2. Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A. 1996. Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J. Biol. Chem. 271: 12687-12690.
    Pubmed CrossRef
  3. Kimberley FC, Screaton GR. 2004. Following a TRAIL: update on a ligand and its five receptors. Cell Res. 14: 359-372.
    Pubmed CrossRef
  4. Malhi H, Gores GJ. 2006. TRAIL resistance results in cancer progression: a TRAIL to perdition? Oncogene 25: 7333-7335.
    Pubmed CrossRef
  5. Hymowitz SG, O’Connell MP, Ultsch MH, Hurst A, Totpal K, Ashkenazi A, et al. 2000. A unique zinc-binding site revealed by a high-resolution X-ray structure of homotrimeric Apo2L/TRAIL. Biochemistry 39: 633-640.
    Pubmed CrossRef
  6. Bodmer JL, Meier P , Tschopp J, Schneider P . 2000. Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL. J. Biol. Chem. 275: 20632-20637.
    Pubmed CrossRef
  7. Szliszka E, Mazur B, Zydowicz G, Czuba ZP, Krol W. 2009. TRAIL-induced apoptosis and expression of death receptor TRAIL-R1 and TRAIL-R2 in bladder cancer cells. Folia Histochem. Cytobiol. 47: 579-585.
    Pubmed
  8. LeBlanc HN, Ashkenazi A. 2003. Apo2L/TRAIL and its death and decoy receptors. Cell Death Differ. 10: 66-75.
    Pubmed CrossRef
  9. Holen I, Croucher PI, Hamdy FC, Eaton CL. 2002. Osteoprotegerin (OPG) is a survival factor for human prostate cancer cells. Cancer Res. 62: 1619-1623.
    Pubmed
  10. Seki N, Hayakawa Y, Brooks AD, Wine J, Wiltrout RH, Yagita H, et al. 2003. Tumor necrosis factor-related apoptosisinducing ligand-mediated apoptosis is an important endogenous mechanism for resistance to liver metastases in murine renal cancer. Cancer Res. 63: 207-213.
    Pubmed
  11. Zhang XD, Nguyen T, Thomas WD, Sanders JE, Hersey P. 2000. Mechanisms of resistance of normal cells to TRAIL induced apoptosis vary between different cell types. FEBS Lett. 482: 193-199.
    CrossRef
  12. Chandrasekaran S, Marshall JR, Messing JA, Hsu JW, King MR. 2014. TRAIL-mediated apoptosis in breast cancer cells cultured as 3D spheroids. PLoS One 9: e111487.
    Pubmed PMC CrossRef
  13. Pan Y, Xu R, Peach M, Huang CP, Branstetter D, Novotny W, et al. 2011. Evaluation of pharmacodynamic biomarkers in a Phase 1a trial of dulanermin (rhApo2L/TRAIL) in patients with advanced tumours. Br. J. Cancer 105: 1830-1838.
    Pubmed PMC CrossRef
  14. Dimberg LY, Anderson CK, Camidge R, Behbakht K, Thorburn A, Ford HL. 2013. On the TRAIL to successful cancer therapy? Predicting and counteracting resistance against TRAIL-based therapeutics. Oncogene 32: 1341-1350.
    Pubmed PMC CrossRef
  15. Stuckey DW, Shah K. 2013. TRAIL on trial: preclinical advances in cancer therapy. Trends Mol. Med. 19: 685-694.
    Pubmed PMC CrossRef
  16. Lim B, Allen JE, Prabhu VV, Talekar MK, Finnberg NK, El-Deiry WS. 2015. Targeting TRAIL in the treatment of cancer: new developments. Expert Opin. Ther. Targets 19:1171-1185.
    Pubmed CrossRef
  17. Schneider P. 2000. Production of recombinant TRAIL and TRAIL receptor: Fc chimeric proteins. Methods Enzymol. 322:325-345.
    CrossRef
  18. Cha SS, Shin HC, Choi KY, Oh BH. 1999. Expression, purification and crystallization of recombinant human TRAIL. Acta Crystallogr. D Biol. Crystallogr. 55: 1101-1104.
    Pubmed CrossRef
  19. Gasparian ME, Ostapchenko VG, Yagolovich AV, Tsygannik IN, Chernyak BV, Dolgikh DA, et al. 2007. Overexpression and refolding of thioredoxin/TRAIL fusion from inclusion bodies and further purification of TRAIL after cleavage by enteropeptidase. Biotechnol. Lett. 29: 1567-1573.
    Pubmed CrossRef
  20. Lin Z, Lei H, Cao P. 2007. Expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosisinducing ligand (TRAIL). Protein Expr. Purif. 51: 276-282.
    Pubmed CrossRef
  21. Wang D, Shi L. 2009. High-level expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Appl. Biochem. Biotechnol. 157: 1-9.
    Pubmed CrossRef
  22. Singh SM, Panda AK. 2005. Solubilization and refolding of bacterial inclusion body proteins. J. Biosci. Bioeng. 99: 303-310.
    Pubmed CrossRef
  23. Li P, Gu Q, Wu X. 2016. Fed-batch production of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in soluble form in Escherichia coli and its purification and characterization. Protein Expr. Purif. 126: 115-121.
    Pubmed CrossRef
  24. Cha SS, Sung BJ, Kim YA, Song YL, Kim HJ, Kim S, et al. 2000. Crystal structure of TRAIL-DR5 complex identifies a critical role of the unique frame insertion in conferring recognition specificity. J. Biol. Chem. 275: 31171-31177.
    Pubmed CrossRef
  25. Vu TT, Koo BK, Song JA, Chong SH, Park CR, Nguyen MT, et al. 2015. Soluble overexpression and purification of bioactive human CCL2 in E. coli by maltose-binding protein. Mol. Biol. Rep. 42: 651-663.
    Pubmed CrossRef
  26. Nguyen MT, Krupa M, Koo BK, Song JA, Vu TT, Do BH, et al. 2016. Prokaryotic soluble overexpression and purification of human VEGF165 by fusion to a maltose binding protein tag. PLoS One 11: e0156296.
    Pubmed PMC CrossRef
  27. Rosano GL, Ceccarelli EA. 2014. Recombinant protein expression in Escherichia coli: advances and challenges. Front. Microbiol. 5: 172.
    Pubmed PMC CrossRef
  28. Vera A, Gonzalez-Montalban N, Aris A, Villaverde A. 2007. The conformational quality of insoluble recombinant proteins is enhanced at low growth temperatures. Biotechnol. Bioeng. 96: 1101-1106.
    Pubmed CrossRef
  29. Schellman JA. 1997. Temperature, stability, and the hydrophobic interaction. Biophys. J. 73: 2960-2964.
    CrossRef
  30. Thiele JR, Habersberger J, Braig D, Schmidt Y, Goerendt K, Maurer V, et al. 2014. Dissociation of pentameric to monomeric C-reactive protein localizes and aggravates inflammation: in vivo proof of a powerful proinflammatory mechanism and a new anti-inflammatory strategy. Circulation 130: 35-50.
    Pubmed CrossRef
  31. Vu TT, Jeong B, Yu J, Koo BK, Jo SH, Robinson RC, et al. 2014. Soluble prokaryotic expression and purification of crotamine using an N-terminal maltose-binding protein tag. Toxicon Official J Int. Soc. Toxinol. 92: 157-165.
  32. Raran-Kurussi S, Waugh DS. 2012. The ability to enhance the solubility of its fusion partners is an intrinsic property of maltose-binding protein but their folding is either spontaneous or chaperone-mediated. PLoS One 7: e49589.
    Pubmed PMC CrossRef
  33. Liu Y, Zhao TJ, Yan YB, Zhou HM. 2005. Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system. Protein Expr. Purif. 44: 155-161.
    Pubmed CrossRef
  34. Nguyen MT, Koo BK, Thi Vu TT, Song JA, Chong SH, Jeong B, et al. 2014. Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase. PLoS One 9: e89038.
    Pubmed PMC CrossRef
  35. Shen YL, Xia XX, Zhang Y, Liu JW, Wei DZ, Yang SL. 2003. Refolding and purification of Apo2l/TRAIL produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli. Biotechnol. Lett. 25: 2097-2101.
    Pubmed CrossRef
  36. de Miguel D, Lemke J, Anel A, Walczak H, Martinez-Lostao L. 2016. Onto better TRAILs for cancer treatment. Cell Death Differ. 23: 733-747.
    Pubmed PMC CrossRef
  37. Trabzuni D, Famulski KS, Ahmad M. 2000. Functional analysis of tumour necrosis factor-alpha-related apoptosisinducing ligand (TRAIL): cysteine-230 plays a critical role in the homotrimerization and biological activity of this novel tumoricidal cytokine. Biochem. J. 350: 505-510.
    Pubmed PMC CrossRef
  38. Gasparian ME, Chernyak BV, Dolgikh DA, Yagolovich AV, Popova EN, Sycheva AM, et al. 2009. Generation of new TRAIL mutants DR5-A and DR5-B with improved selectivity to death receptor 5. Apoptosis 14: 778-787.
    Pubmed CrossRef
  39. Li JH, Kirkiles-Smith NC, McNiff JM, Pober JS. 2003. TRAIL induces apoptosis and inflammatory gene expression in human endothelial cells. J. Immunol. 171: 1526-1533.
    Pubmed CrossRef
  40. Qiu F, Hu M, Tang B, Liu X, Zhuang H, Yang J, et al. 2013. Annexin V-TRAIL fusion protein is a more sensitive and potent apoptotic inducer for cancer therapy. Sci. Rep. 3: 3565.
    Pubmed PMC CrossRef
  41. Cao L, Du P, Jiang SH, Jin GH, Huang QL, Hua ZC. 2008. Enhancement of antitumor properties of TRAIL by targeted delivery to the tumor neovasculature. Mol. Cancer Ther. 7:851-861.
    Pubmed CrossRef
  42. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, et al. 1995. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3: 673-682.
    CrossRef
  43. Kim SH, Kim K, Kwagh JG, Dicker DT, Herlyn M, Rustgi AK, et al. 2004. Death induction by recombinant native TRAIL and its prevention by a caspase 9 inhibitor in primary human esophageal epithelial cells. J. Biol. Chem. 279: 4004440052.
    CrossRef
  44. van der Sloot AM, Mullally MM, Fernandez-Ballester G, Serrano L, Quax WJ. 2004. Stabilization of TRAIL, an allbetasheet multimeric protein, using computational redesign. Protein Eng. Des. Sel. 17: 673-680.
    Pubmed CrossRef
  45. Pan LQ, Zhao WB, Lai J, Ding D, Wei XY, Li YY, et al. 2015. Hetero-modification of TRAIL trimer for improved drug delivery and in vivo antitumor activities. Sci. Rep. 5: 14872.
    Pubmed PMC CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2017; 27(12): 2156-2164

Published online December 28, 2017 https://doi.org/10.4014/jmb.1705.05070

Copyright © The Korean Society for Microbiology and Biotechnology.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand

Bich Hang Do 1, Minh Tan Nguyen 1, Jung-A Song 1, Sangsu Park 1, Jiwon Yoo 1, Jaepyeong Jang 1, Sunju Lee 1, Seoungjun So 1, Yejin Yoon 1, Inki Kim 2, Kyungjin Lee 2, Yeon Jin Jang and Han Choe 1*

1Department of Physiology, Asan-Minnesota Institute for Innovating Transplantation, Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, Republic of Korea, 2Department of Convergence Medicine, Asan Institute for Life Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, Republic of Korea, 3Department of Physiology, Cell Dysfunction Research Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea

Received: May 25, 2017; Accepted: September 25, 2017

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an
antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting
toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble
expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL
by fusing maltose-binding protein (MBP), b’a’ domain of protein disulfide isomerase
(PDIb’a’), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified
using subsequent immobilized metal affinity chromatography and amylose-binding
chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5
mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%.
The endotoxin level of the final product was 0.4 EU/μg, as measured by the Limulus
amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause
apoptosis of HeLa cells with an EC50 and Hill coefficient of 0.6 ± 0.03 nM and 2.41 ± 0.15,
respectively. The high level of apoptosis in HeLa cells following administration of purified
TRAIL indicates the significance and novelty of this method for producing high-grade and
high-yield TRAIL.

Keywords: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), expression, purification, E. coli, cytotoxic activity, HeLa cells

References

  1. Wang S, El-Deiry WS. 2003. TRAIL and apoptosis induction by TNF-family death receptors. Oncogene 22: 8628-8633.
    Pubmed CrossRef
  2. Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A. 1996. Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J. Biol. Chem. 271: 12687-12690.
    Pubmed CrossRef
  3. Kimberley FC, Screaton GR. 2004. Following a TRAIL: update on a ligand and its five receptors. Cell Res. 14: 359-372.
    Pubmed CrossRef
  4. Malhi H, Gores GJ. 2006. TRAIL resistance results in cancer progression: a TRAIL to perdition? Oncogene 25: 7333-7335.
    Pubmed CrossRef
  5. Hymowitz SG, O’Connell MP, Ultsch MH, Hurst A, Totpal K, Ashkenazi A, et al. 2000. A unique zinc-binding site revealed by a high-resolution X-ray structure of homotrimeric Apo2L/TRAIL. Biochemistry 39: 633-640.
    Pubmed CrossRef
  6. Bodmer JL, Meier P , Tschopp J, Schneider P . 2000. Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL. J. Biol. Chem. 275: 20632-20637.
    Pubmed CrossRef
  7. Szliszka E, Mazur B, Zydowicz G, Czuba ZP, Krol W. 2009. TRAIL-induced apoptosis and expression of death receptor TRAIL-R1 and TRAIL-R2 in bladder cancer cells. Folia Histochem. Cytobiol. 47: 579-585.
    Pubmed
  8. LeBlanc HN, Ashkenazi A. 2003. Apo2L/TRAIL and its death and decoy receptors. Cell Death Differ. 10: 66-75.
    Pubmed CrossRef
  9. Holen I, Croucher PI, Hamdy FC, Eaton CL. 2002. Osteoprotegerin (OPG) is a survival factor for human prostate cancer cells. Cancer Res. 62: 1619-1623.
    Pubmed
  10. Seki N, Hayakawa Y, Brooks AD, Wine J, Wiltrout RH, Yagita H, et al. 2003. Tumor necrosis factor-related apoptosisinducing ligand-mediated apoptosis is an important endogenous mechanism for resistance to liver metastases in murine renal cancer. Cancer Res. 63: 207-213.
    Pubmed
  11. Zhang XD, Nguyen T, Thomas WD, Sanders JE, Hersey P. 2000. Mechanisms of resistance of normal cells to TRAIL induced apoptosis vary between different cell types. FEBS Lett. 482: 193-199.
    CrossRef
  12. Chandrasekaran S, Marshall JR, Messing JA, Hsu JW, King MR. 2014. TRAIL-mediated apoptosis in breast cancer cells cultured as 3D spheroids. PLoS One 9: e111487.
    Pubmed KoreaMed CrossRef
  13. Pan Y, Xu R, Peach M, Huang CP, Branstetter D, Novotny W, et al. 2011. Evaluation of pharmacodynamic biomarkers in a Phase 1a trial of dulanermin (rhApo2L/TRAIL) in patients with advanced tumours. Br. J. Cancer 105: 1830-1838.
    Pubmed KoreaMed CrossRef
  14. Dimberg LY, Anderson CK, Camidge R, Behbakht K, Thorburn A, Ford HL. 2013. On the TRAIL to successful cancer therapy? Predicting and counteracting resistance against TRAIL-based therapeutics. Oncogene 32: 1341-1350.
    Pubmed KoreaMed CrossRef
  15. Stuckey DW, Shah K. 2013. TRAIL on trial: preclinical advances in cancer therapy. Trends Mol. Med. 19: 685-694.
    Pubmed KoreaMed CrossRef
  16. Lim B, Allen JE, Prabhu VV, Talekar MK, Finnberg NK, El-Deiry WS. 2015. Targeting TRAIL in the treatment of cancer: new developments. Expert Opin. Ther. Targets 19:1171-1185.
    Pubmed CrossRef
  17. Schneider P. 2000. Production of recombinant TRAIL and TRAIL receptor: Fc chimeric proteins. Methods Enzymol. 322:325-345.
    CrossRef
  18. Cha SS, Shin HC, Choi KY, Oh BH. 1999. Expression, purification and crystallization of recombinant human TRAIL. Acta Crystallogr. D Biol. Crystallogr. 55: 1101-1104.
    Pubmed CrossRef
  19. Gasparian ME, Ostapchenko VG, Yagolovich AV, Tsygannik IN, Chernyak BV, Dolgikh DA, et al. 2007. Overexpression and refolding of thioredoxin/TRAIL fusion from inclusion bodies and further purification of TRAIL after cleavage by enteropeptidase. Biotechnol. Lett. 29: 1567-1573.
    Pubmed CrossRef
  20. Lin Z, Lei H, Cao P. 2007. Expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosisinducing ligand (TRAIL). Protein Expr. Purif. 51: 276-282.
    Pubmed CrossRef
  21. Wang D, Shi L. 2009. High-level expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Appl. Biochem. Biotechnol. 157: 1-9.
    Pubmed CrossRef
  22. Singh SM, Panda AK. 2005. Solubilization and refolding of bacterial inclusion body proteins. J. Biosci. Bioeng. 99: 303-310.
    Pubmed CrossRef
  23. Li P, Gu Q, Wu X. 2016. Fed-batch production of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in soluble form in Escherichia coli and its purification and characterization. Protein Expr. Purif. 126: 115-121.
    Pubmed CrossRef
  24. Cha SS, Sung BJ, Kim YA, Song YL, Kim HJ, Kim S, et al. 2000. Crystal structure of TRAIL-DR5 complex identifies a critical role of the unique frame insertion in conferring recognition specificity. J. Biol. Chem. 275: 31171-31177.
    Pubmed CrossRef
  25. Vu TT, Koo BK, Song JA, Chong SH, Park CR, Nguyen MT, et al. 2015. Soluble overexpression and purification of bioactive human CCL2 in E. coli by maltose-binding protein. Mol. Biol. Rep. 42: 651-663.
    Pubmed CrossRef
  26. Nguyen MT, Krupa M, Koo BK, Song JA, Vu TT, Do BH, et al. 2016. Prokaryotic soluble overexpression and purification of human VEGF165 by fusion to a maltose binding protein tag. PLoS One 11: e0156296.
    Pubmed KoreaMed CrossRef
  27. Rosano GL, Ceccarelli EA. 2014. Recombinant protein expression in Escherichia coli: advances and challenges. Front. Microbiol. 5: 172.
    Pubmed KoreaMed CrossRef
  28. Vera A, Gonzalez-Montalban N, Aris A, Villaverde A. 2007. The conformational quality of insoluble recombinant proteins is enhanced at low growth temperatures. Biotechnol. Bioeng. 96: 1101-1106.
    Pubmed CrossRef
  29. Schellman JA. 1997. Temperature, stability, and the hydrophobic interaction. Biophys. J. 73: 2960-2964.
    CrossRef
  30. Thiele JR, Habersberger J, Braig D, Schmidt Y, Goerendt K, Maurer V, et al. 2014. Dissociation of pentameric to monomeric C-reactive protein localizes and aggravates inflammation: in vivo proof of a powerful proinflammatory mechanism and a new anti-inflammatory strategy. Circulation 130: 35-50.
    Pubmed CrossRef
  31. Vu TT, Jeong B, Yu J, Koo BK, Jo SH, Robinson RC, et al. 2014. Soluble prokaryotic expression and purification of crotamine using an N-terminal maltose-binding protein tag. Toxicon Official J Int. Soc. Toxinol. 92: 157-165.
  32. Raran-Kurussi S, Waugh DS. 2012. The ability to enhance the solubility of its fusion partners is an intrinsic property of maltose-binding protein but their folding is either spontaneous or chaperone-mediated. PLoS One 7: e49589.
    Pubmed KoreaMed CrossRef
  33. Liu Y, Zhao TJ, Yan YB, Zhou HM. 2005. Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system. Protein Expr. Purif. 44: 155-161.
    Pubmed CrossRef
  34. Nguyen MT, Koo BK, Thi Vu TT, Song JA, Chong SH, Jeong B, et al. 2014. Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase. PLoS One 9: e89038.
    Pubmed KoreaMed CrossRef
  35. Shen YL, Xia XX, Zhang Y, Liu JW, Wei DZ, Yang SL. 2003. Refolding and purification of Apo2l/TRAIL produced as inclusion bodies in high-cell-density cultures of recombinant Escherichia coli. Biotechnol. Lett. 25: 2097-2101.
    Pubmed CrossRef
  36. de Miguel D, Lemke J, Anel A, Walczak H, Martinez-Lostao L. 2016. Onto better TRAILs for cancer treatment. Cell Death Differ. 23: 733-747.
    Pubmed KoreaMed CrossRef
  37. Trabzuni D, Famulski KS, Ahmad M. 2000. Functional analysis of tumour necrosis factor-alpha-related apoptosisinducing ligand (TRAIL): cysteine-230 plays a critical role in the homotrimerization and biological activity of this novel tumoricidal cytokine. Biochem. J. 350: 505-510.
    Pubmed KoreaMed CrossRef
  38. Gasparian ME, Chernyak BV, Dolgikh DA, Yagolovich AV, Popova EN, Sycheva AM, et al. 2009. Generation of new TRAIL mutants DR5-A and DR5-B with improved selectivity to death receptor 5. Apoptosis 14: 778-787.
    Pubmed CrossRef
  39. Li JH, Kirkiles-Smith NC, McNiff JM, Pober JS. 2003. TRAIL induces apoptosis and inflammatory gene expression in human endothelial cells. J. Immunol. 171: 1526-1533.
    Pubmed CrossRef
  40. Qiu F, Hu M, Tang B, Liu X, Zhuang H, Yang J, et al. 2013. Annexin V-TRAIL fusion protein is a more sensitive and potent apoptotic inducer for cancer therapy. Sci. Rep. 3: 3565.
    Pubmed KoreaMed CrossRef
  41. Cao L, Du P, Jiang SH, Jin GH, Huang QL, Hua ZC. 2008. Enhancement of antitumor properties of TRAIL by targeted delivery to the tumor neovasculature. Mol. Cancer Ther. 7:851-861.
    Pubmed CrossRef
  42. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, et al. 1995. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3: 673-682.
    CrossRef
  43. Kim SH, Kim K, Kwagh JG, Dicker DT, Herlyn M, Rustgi AK, et al. 2004. Death induction by recombinant native TRAIL and its prevention by a caspase 9 inhibitor in primary human esophageal epithelial cells. J. Biol. Chem. 279: 4004440052.
    CrossRef
  44. van der Sloot AM, Mullally MM, Fernandez-Ballester G, Serrano L, Quax WJ. 2004. Stabilization of TRAIL, an allbetasheet multimeric protein, using computational redesign. Protein Eng. Des. Sel. 17: 673-680.
    Pubmed CrossRef
  45. Pan LQ, Zhao WB, Lai J, Ding D, Wei XY, Li YY, et al. 2015. Hetero-modification of TRAIL trimer for improved drug delivery and in vivo antitumor activities. Sci. Rep. 5: 14872.
    Pubmed KoreaMed CrossRef