Amidolytic Activity of Factor VII Expressed in Iranian Lizard Leishmania
Trends in Peptide and Protein Sciences,
Vol. 3 (2018),
1 June 2018
,
Page 1-5 (e2)
https://doi.org/10.22037/tpps.v3i0.20924
Abstract
Over many years, variable gene expression systems have been used obtaining of Factor VII protein, but each one has certain limitations. Therefore, the main goal of this study was to assess the biological activity of purified and recombinant factor VII expressed in Iranian lizard Leishmania. After transferring recombinant construct containing FVII gene to Leishmania, first, the expression of 55 kDa FVII protein in transfected cells was confirmed by analyzing cell lysate using SDS-PAGE and Western Blotting techniques. Then, FVII was purified by NI-NTA-His Tag- resin through an affinity chromatography. The chromogenic activity of human recombinant Factor VII (amidolytic activity) in supernatant and pellet fractions of Leishmania promastigotes was done using ELISA method. The amidolytic activity of rhFVII was about 0.0125 IU/ml in the concentrated cell sediment and 0 IU/ml in the supernatant in the first 60 minutes and after that time, none of the samples showed acceptable activity.
HIGHLIGHTS
• Factor VII has important role in the treatment of coagulation and hemophilic disease.
• Recombinant FVII was expressed in Iranian Lizard Leishmania.
• The amidolytic activity of rhFVII was about 0.0125 IU/ml in cell pellet.
• No amildolytic activity was observed in the supernatant.
- Amidolytic activity
- Chromogenic assay
- Lizard Leishmania
- Recombinant factor VII
How to Cite
References
Basile, G. and M. Peticca, (2009). “Recombinant protein expression in Leishmania tarentolae.” Molecular Biotechnology, 43(3): 273.
Davie, E. W., Fujikawa, K., Kurachi, K. and W. Kisiel, (1979). “The role of serine proteases in the blood coagulation cascade.” Adv Enzymol Relat Areas Mol Biol, 48: 277-318.
Fritsche, C., Sitz, M., Weiland, N., Breitling, R. and H. D. Pohl, (2007). “Characterization of the growth behavior of Leishmania tarentolae–a new expression system for recombinant proteins.” Journal of Basic Microbiology, 47(5): 384-393.
Fritsche, C., Sitz, M., Wolf, M. and H. D. Pohl, (2008). “Development of a defined medium for heterologous expression in Leishmania tarentolae.” Journal of Basic Microbiology, 48(6): 488-495.
Hagen, F. S., Murray, M. J., Busby, S. J., Berkner, K. L., Insley, M. Y., Woodbury, R. G. and C. L. Gray, (1988). ″Expression of factor VII activity in mammalian cells″, Google Patents.
Halabian, R., Roudkenar, M. H., Esmaeili, N. S., Masroori, N., Roushandeh, A. and A. Najafabadi, (2009). “Establishment of a cell line expressing recombinant factor VII and its subsequent conversion to active form FVIIa through hepsin by genetic engineering method.” Vox Sanguinis, 96(4): 309-315.
Hedner, U. and W. Kisiel, (1983). “Use of human factor VIIa in the treatment of two hemophilia A patients with high-titer inhibitors.” The Journal of Clinical Investigation, 71(6): 1836-1841.
Hoffman, C. J., Miller, R. H., Lawson, W. E. and M. B. Hultin, (1989). “Elevation of factor VII activity and mass in young adults at risk of ischemic heart disease.” Journal of the American College of Cardiology, 14(4): 941-946.
Hoffman, M., Monroe III, D. and H. Roberts, (1998). “Activated factor VII activates factors IX and X on the surface of activated platelets: thoughts.” Blood Coagulation and Fibrinolysis, 9(1): S61-S65.
Kazemi, B., Tahvidar- Bideroni, G., Hashemi Feshareki, S.R. and E. Javadian (2004). “Isolation a lizard Leishmania promastigote from natural host in Iran.” Journal of Biological Sciences, 4(5): 620-623.
Khan, A. H., Bayat, H., Rajabibazl, M., Sabri, S. and A. Rahimpour, (2017). “Humanizing glycosylation pathways in eukaryotic expression systems.” World Journal of Microbiology and Biotechnology, 33(1): 4.
Kubisz, P., Stasko, J., Holley, P. (2006). “Recombinant technology in hemostatic disorders.” In: Wiwanitkit, V. (ed). Thrombohemistatic Disease Research. New York: Nova Science Publisher, p. 47.
Masroori, N., Halabian, R., Mohammadipour, M., Roushandeh, A. M., Rouhbakhsh, M., Najafabadi, A. J., Fathabad, M. E., Salimi, M., Shokrgozar, M. A. and M. H. Roudkenar, (2010). “High-level expression of functional recombinant human coagulation factor VII in insect cells.” Biotechnology Letters, 32(6):803-809.
Mirzaahmadi, S., Asaadi-Tehrani, G., Bandehpour, M., Davoudi, N., Tahmasbi, L., Hosseinzadeh, N., Mirzahoseini, H., Parivar, K. and B. Kazemi ,(2011). “Expression of recombinant human coagulation factor VII by the Lizard Leishmania expression system. " BioMed Research International, 2011.
Niimi, T., (2012). “Recombinant protein production in the eukaryotic protozoan parasite Leishmania tarentolae: a review”. Recombinant Gene Expression, Springer: 307-315.
Østerud, B., (1983). “How to measure factor VII and factor VII activation.” Pathophysiology of Haemostasis and Thrombosis, 13(3): 161-168.
Pedersen, A. H., Lund-Hansen, T., Bisgaard-Frantzen, H., Olsen, F. and L. C. Petersen, (1989). “Autoactivation of human recombinant coagulation factor VII.” Biochemistry, 28(24):9331-9336.
Persson, E., Olsen, O. H., Østergaard, A. and L. S. Nielsen, (1997). “Ca2+ binding to the first epidermal growth factor-like domain of factor VIIa increases amidolytic activity and tissue factor affinity.” Journal of Biological Chemistry, 272(32):19919-19924.
Robinson, K. A. and S. M. Beverley, (2003). “Improvements in transfection efficiency and tests of RNA interference (RNAi) approaches in the protozoan parasite Leishmania.” Molecular and Biochemical Parasitology, 128(2): 217-228.
Seligsohn ,U., Osterud, B. and S. I. Rapaport, (1978). “Coupled amidolytic assay for factor VII: its use with a clotting assay to determine the activity state of factor VII.” Blood, 52(5): 978-988.
Sodoyer, R., (2004). “Expression systems for the production of recombinant pharmaceuticals.” BioDrugs, 18(1): 51-62.
Suzuki, T., Yamauchi, K., Matsushita, T., Furumichi, T., Furui, H., Tsuzuki, J. and H. Saito (1991). “Elevation of factor VII activity and mass in coronary artery disease of varying severity.” Clinical Cardiology, 1(9)4:731-736.
van Dieijen-Visser, M., Van Wersch, J., Brombacher, P., Rosing, J., Hemker, H. and G. Van Dieijen (1982). “Use of chromogenic peptide substrates in the determination of clotting factors II, VII, IX and X in normal plasma and in plasma of patients treated with oral anticoagulants.” Pathophysiology of Haemostasis and Thrombosis, 12(3): 241-255.
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