Abstract
The modified asparaginase Was79 was derived from the recombinant wild-type l-asparaginase of Wolinella succinogenes. The Was79 contains the amino acid substitutions V23Q and K24T responsible for the resistance to trypsinolysis and the N-terminal heparin-binding peptide KRKKKGKGLGKKR responsible for the binding to heparin and tumor K562 cells in vitro. When tested on a mouse model of Fischer lymphadenosis L5178Y, therapeutic efficacy of Was79 was significantly higher than that of reference enzymes at all single therapeutic doses used (125–8000 IU/kg). At Was79 single doses of 500–8000 IU/kg, the complete remission rate of 100 % was observed. The Was79 variant can be expressed intracellularly in E. coli as a less immunogenic formyl-methionine-free form at high per cell production levels.
Similar content being viewed by others
References
Durden, D. L., & Distasio, J. A. (1980). Comparison of the immunosuppressive effects of asparaginases from Escherichia coli and Vibrio succinogenes. Cancer Research, 40, 1125–1129.
Durden, D. L., & Distasio, J. A. (1981). Characterization of the effects of asparaginases from Escherichia coli and a glutaminase free asparaginase from Vibrio succinogenes on specific cell-mediated cytotoxicity. International Journal of Cancer, 27, 59–65.
van den Berg, H. (2011). Asparaginase revisited. Leukemia and Lymphoma, 52, 168–178.
Covini, D., Tardito, S., Bussolati, O., Laurent, R., Chiarelli, L. R., Pasquetto, M. V., et al. (2012). Expanding targets for a metabolic therapy of cancer: l-Asparaginase. Recent Patents on Anti-Cancer Drug Discovery, 7, 4–13.
Shrivastava, A., Khan, A. A., Khurshid, M., Kalam, M. A., Jain, S. K., & Singhal, P. K. (2015). Recent developments in l-asparaginase discovery and its potential as anticancer agent. Critical Reviews in Oncology/Hematology, 100, 1–10.
Distasio, J. A., Niederman, R. A., Kafkewitz, D., & Goodman, D. (1976). Purification and characterization of l-asparaginase with anti-lymphoma activity from Vibrio succinogenes. Journal of Biological Chemistry, 251, 6929–6933.
Distasio, J. A., Salazar, A. M., Nadji, M., & Durden, D. L. (1982). Glutaminase-free asparaginase from Vibrio succinogenes: an antilymphoma enzyme lacking hepatotoxicity. International Journal of Cancer, 30, 343–347.
Lubkowski, J., Palm, G. J., Gilliland, G. L., Derst, C., Röhm, K.-H., & Wlodawer, A. (1996). Crystal structure and amino acid sequence of Wolinella succinogenes l-asparaginase. European Journal of Biochemistry, 241, 201–207.
Distasio, J. A., Niederman, R. A., & Kafkewitz, D. (1977). Antilymphoma activity of a glutaminase-free L-asparaginase of microbial origin. Proceedings of the Society for Experimental Biology and Medicine, 155, 528–531.
Reinert, R. B., Oberle, L. M., Wek, S. A., Bunpo, P., Wang, X. P., Mileva, I., et al. (2006). Role of glutamine depletion in directing tissue-specific nutrient stress responses to l-asparaginase. Journal of Biological Chemistry, 281, 31222–31233.
Derst, C., Henseling, J., & Röhm, K.-H. (2000). Engineering the substrate specificity of Escherichia coli asparaginase II. Selective reduction of glutaminase activity by amino acid replacements at position 248. Protein Science, 9, 2009–2017.
Kotzia, G. A., Lappa, K., & Labrou, N. E. (2007). Tailoring structure-function properties of l-asparaginase: engineering resistance to trypsin cleavage. Biochemical Journal, 404, 337–343.
Xu, D., & Esko, J. D. (2014). Demystifying heparan sulfate: Protein interactions. Annual Review of Biochemistry, 83, 129–157.
Thompson, S. A., Higashiyama, S., Wood, K., Pollitt, N. S., Damm, D., McEnroe, G., et al. (1994). Characterization of sequences within heparin-binding EGF-like growth factor that mediate interaction with heparin. Journal of Biological Chemistry, 269, 2541–2549.
Neubauer, P., Hofmann, K., Holst, O., Mattiasson, B., & Kruschke, P. (1992). Maximizing the expression of a recombinant gene in Escherichia coli by manipulation of induction time using lactose as inducer. Applied Microbiology and Biotechnology, 36, 739–744.
Studier, F. W. (2005). Protein production by auto-induction in high-density shaking cultures. Protein Expression and Purification, 41, 207–234.
Wriston, J. C. (1970). Asparaginase. Methods in Enzymology, 17A, 732–742.
Dawson, R. M. C., Elliott, D. C., Elliott, W. H., & Jones, K. M. (1986). Data for biochemical research (3rd ed.). Oxford: Clarendon Press.
The, T. H., & Feltkamp, T. E. W. (1970). Conjugation of fluorescein isothiocyanate to antibodies. I. Experiments on the conditions of conjugation. Immunology, 18, 865–873.
Chabner, B. A., & Longo, D. L. (Eds.). (2001). Cancer chemotherapy and biotherapy: Principles and practice (3rd ed.). Philadelphia: Lippincott Williams & Wilkins.
Wade, H. E., Elsworth, R., Herbert, D., Keppie, J., & Sargeant, K. (1968). A new l-asparaginase with antitumor activity. Lancet, 2, 776–777.
Rabiet, M. J., Huet, E., & Boulay, F. (2007). The N-formyl peptide receptors and the anaphylatoxin C5a receptors: an overview. Biochimie, 89, 1089–1106.
Elleuche, S., & Pöggeler, S. (2010). Inteins, valuable genetic elements in molecular biology and biotechnology. Applied Microbiology and Biotechnology, 87, 479–489.
Burchenal, J. H., Benvenisti, D., & Dollinger, M. (1970). Experimental studies with l-asparaginase in mouse leukemias. In E. Grundmann & H. F. Oettgen (Eds.), Experimental and clinical effects of l -asparaginase (Vol. 33, pp. 102–113)., Recent results in cancer research Berlin: Springer.
Connors, T. A., & Jones, M. (1970). The effect of asparaginase on some animal-tumors. In E. Grundmann & H. F. Oettgen (Eds.), Experimental and clinical effects of l -asparaginase (Vol. 33, pp. 181–187)., Recent results in cancer research Berlin: Springer.
Offman, M. N., Krol, M., Patel, N., Krishnan, S., Liu, J.-Z., Saha, V., & Bates, P. A. (2011). Rational engineering of l-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood, 117, 1614–1621.
Chan, W. K., Lorenzi, P. L., Anishkin, A., Purwaha, P., Rogers, D. M., Sukharev, S., et al. (2014). The glutaminase activity of l-asparaginase is not required for anticancer activity against ASNS-negative cells. Blood, 123, 3596–3606.
Wriston, J. C, Jr, & Yellin, T. O. (1973). l-asparaginase: A review. Advances in Enzymology and Related Areas of Molecular Biology, 39, 185–248.
Schweizer, F. (2009). Cationic amphiphilic peptides with cancer-selective toxicity. European Journal of Pharmacology, 625, 190–194.
Kwon, Y. M., Chung, H. S., Moon, C., Yockman, J., Park, Y. J., Gitlin, S. D., et al. (2009). l-Asparaginase-encasulated intact erythrocytes for treatment of acute lymphoblastic leukemia (ALL). Journal of Controlled Release, 139, 182–189.
Domenech, C., Thomas, X., Chabaud, S., Baruchel, A., Gueyffier, F., Mazingue, F., et al. (2011). l-Asparaginase loaded red blood cells in refractory or relapsing acute lymphoblastic leukaemia in children and adults: results of the GRASPALL 2005-01 randomized trial. British Journal of Haematology, 153, 58–65.
Agrawal, V., Woo, J. H., Borthakur, G., Kantarjian, H., & Frankel, A. E. (2013). Red blood cell-encapsulated l-asparaginase: potential therapy of patients with asparagine synthetase deficient acute myeloid leukemia. Protein and Peptide Letters, 20, 392–402.
Lorentz, C. M., Kontos, S., Diaceri, G., Henry, H., & Hubbell, J. A. (2015). Engineered binding to erythrocytes induces immunological tolerance to E. coli asparaginase. Science Advances, 1, e1500112.
Acknowledgments
This work was supported by the Ministry of Education and Science of RF (State contract No. 14.N08.11.0014).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflicts of interest.
Ethical approval
All applicable international, national, and institutional guidelines for the care and use of animals were followed.
Rights and permissions
About this article
Cite this article
Sannikova, E.P., Bulushova, N.V., Cheperegin, S.E. et al. The Modified Heparin-Binding l-Asparaginase of Wolinella succinogenes . Mol Biotechnol 58, 528–539 (2016). https://doi.org/10.1007/s12033-016-9950-1
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12033-016-9950-1