Fibrinolytic Activity of Recombinant Mutant Streptokinase
Novelty in Biomedicine,
Vol. 3 No. 2 (2015),
12 April 2015
,
Page 89-93
https://doi.org/10.22037/nbm.v3i2.8679
Abstract
Background: Streptokinase is a bacterial protein produced by different beta hemolytic streptococci and widely used in thrombolytic treatment. The main disadvantage of using streptokinase is antibody formation which causes allergic reaction to neutralize effects of streptokinase therapy. Aim of this study was investigate of recombinant mutant streptokinase fibrinolytic activity.Materials and Methods: In this study recombinant mutant streptokinase without 42 amino acids from the C terminal region was purified by affinity S-Tag column chromatography and its fibrinolytic activity was studied.
Results: The concentration of expressed and purified protein was 10 mg/ml. Its enzyme activity was assayed using zymography, radial caseinolytic activity and fibrin plate test methods and estimated quantitatively by casein digestion method compared to a commercial form.
Conclusion: It was found that this product had the more volume and more enzymatic activity.
- Mutant streptokinase
- Zymography
- Caseinolytic activity
How to Cite
References
Stehle G, Schettler G. Review and current status of thrombolytic therapy with streptokinase. Tokai J Exp Clin Med. 1986;11(1):51-66.
Collen D. Coronary thrombolysis: streptokinase or recombinant tissue-type plasminogen activator? Ann Intern Med. 1990;112: 529–38.
Banerjee AY, Chisti UC, Banerjee A. Streptokinase clinically useful thrombolytic agent. Biotechnol Adv. 2004;22:287-307.
De Renzo EC, Siiteri PK, Hutchings BL, Bell PH. Preparation and certain properties of highly purified streptokinase. J Biol Chem. 1967;242:533–42.
Taylor FB, Botts J. Purification and characterization of streptokinase with studies of streptokinase activation of plasminogen. Biochemistry. 1968;7:232–42.
Brockway WJ, Castellino FJ. A characterization of native streptokinase and altered streptokinase isolated from a human plasminogen activator complex. Biochemistry 1974; 13: 2063– 2070.
McCoy HE, Broder CC, Lottenberg R. Streptokinases produced by pathogenic group C Streptococci demostrate species specific plasminogen activation. J Infect Dis. 1991;164:515-21.
Welfle K, Pfeil W, Misselwitz R, Welfle H, Gerlach D. Conformational properties of streptokinase-differential scanning calorimetric investigations. Int J Biol Macromol 1992; 14: 19-22.
Loy JA, Lin X, Schenone M, Castellino FJ, Zhang XC, Tang J. Domain interactions between streptokinase and human plasminogen. Biochemistry. 2001;40(48):14686-95.
Nihalani D, Kumar R, Rajagopal K, Sahni G. Role of the amino-terminal region of streptokinase in the generation of a fully functional plasminogen activator complex probed with synthetic peptides. Prot Sci. 1998;7:637-48.
Kim DM, Lee SJ, Yoon SK, Byun SM. Specificity role of the streptokinase terminal domain in plasminogen activation. Biochem Biophys Res Commun. 2002;290(1):585-8.
Zhai P, Wakeham N, Loy JA, Zhang XC. Functional roles of streptokinase C terminal flexible peptide in active site formation and substrate recognition in plasminogen activation. Biochemistry. 2003;42:114-20.
Kim DM, Lee SJ, Kim IC, Kim ST, Byun SM. Asp4-His48 region of streptokinase is important in binding to a substrate plasminogen. Thromb Res. 2000;99:93-8.
Tillet WS, Edwards LB, Garner RL. Fibrinolytic activity of haemolytic streptococci: Development of resistance of fibrinolysis following acute haemolytic streptococcus infection. J Clin Invest. 1934;13:47-78.
Brugemann J, Bom VJJ. Anti-streptokinase antibodies inhibit fibrinolytic effects of Antistreptokinase in acute myocardial infarction. Am J Cardiol. 1993;72:462-4.
Reed GL, Kussie P, Parhami–seren B. A functional analysis of the antigenicity of streptokinase using monoclonal antibody mapping and recombinant streptokinase fragments. J Immunol. 1993;150:4407-15.
Parhami–Seren B, Harvey ML, White D, Reed GL. Mapping the antigenic region of streptokinase in humans before and after streptokinase therapy. Mol Immunol. 1995;32:717-24.
Ojalvo AG, Pozo L, Labarta V, Torrens I. Prevalence of circulating antibodies against streptokinase C-terminal peptide in normal blood donors. Biochem. Biophys. Res. Commun. 1999;263(2):454-9.
Torrens I, Reyes O, Ojalvo AG, Seralena A, Chinea G, Cruz LJ, de la Fuente J. Mapping of the antigenic regions of streptokinase in humans after streptokinase therapy. Biochem Biophys Res Commun. 1999;259:162–8.
Bandehpour M, Sharifnia Z, Mohajeri N, Taherkhani M, Koochaki A, Yarian F, et al. Comparative study of the reactivity of natural and mutated streptokinase with total antistreptokinase antibodies in human sera. Blood Coagul. Fibrinolysis. 2012;23(8):734-8.
Kazemi B, Fallahian F, Seyed N, Bandehpour M, Shabani A, Ghadam P. Molecular cloning of the streptokinase mutant gene. Pakis JBiol Sci. 2006;9:557–9.
Seyed N, Shekari P, Bandehpour, M, Sharifnia Z, Parivar K, Kazemi B. Production of mutant streptokinase recombinant protein. Iran. J Clin Infect Dis. 2008;3:179–83.
Shukla AA, Peterson J, Sorge L, Lewis P, Thomas S, Waugh S. Preparative purification of a recombinant protein by hydrophobic interaction chromatography: modulation of selectivity by the use of chaotropic additives. Biotechnol Prog. 2002;18(3):556-64.
Kim YS, Jones LS, Dong A, Kendrick BS, Chang BS, Manning MC, et al. Effects of sucrose on conformational equilibria and fluctuations within the native-state ensemble of proteins. Prot Sci. 2003;12:1252–61.
Bischoff KM, Shi L, Kennelly PJ. The detection of enzyme activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Anal. Biochem. 1998;260(1):1-17.
Lantz MS, Ciborowski P. Zymographic techniques for detection and characterization of microbial proteases. Methods Enzymol. 1994;235:563-94.
Palolahti M, Lauharanta J, Stephens RW, Kuusela P, Vaheri A. Proteolytic activity in leg ulcer exudate. Exp Dermatol. 1993;2(1):29-37.
Astrup T, Mullertz S. The fibrin plate method for estimating fibrinolytic activity. Arch. Biochem. Biophy. 1952;40:346–51.
Mounter LA, Shipley BA. The inhibition of plasmin by toxic phosphorus compounds. J Biol Chem. 1957;226:855-61.
Christensen LR. Streptococcal fibrinolysis: a proteolytic reaction due to a serum enzyme activated by streptococcal fibrinolysin. J Gen Physiol. 1944;28:363-83.
Norman PS. Studies of the plasmin system, i. measiyrement of human and animam plasminogen. Measurement of an activator in Human serum. J Exp Med. 1957;106(3):423-37.
Markus G, Werkheiser WC. The Interaction of Streptokinase with Plasminogen. 1 Functional Properties of The Activated Enzyme. J Biol Chem. 1964;239:2637-43.
Baruah DB, Dash RN, Chaundhari MR, Kadam SS. Plasminogen Activators: A Comparison. Vascul Pharmacol. 2004;44:1-9.
Zhang Y, Liang Z, Hsueh HT, Ploplis VA, Castellino FJ. Characterization of streptokinases from group A streptococci reveals a strong functional relationship that supports the coinheritance of plasminogen-binding M-protein and cluster 2B streptokinase. J Biol Chem. 2012;287(50):42093-103 .
Jackson KW, Tang J. Complete amino acid sequence of streptokinase and its homology with serine proteases. Biochemistry. 1982;21(26):6620-5.
Boxrud PD, Bock PE. Streptokinase binds preferentially to the extended conformation of plasminogen through lysine binding site and catalytic domain interactions. Biochemistry. 2000;39(45):13974-81.
Wang X, Lin X, Lay JA, Tang J. Zhang XC. Crystal structure of the catalytic domain of human plasmin complexed with streptokinase. Science. 1998;281:1662–5.
Torrens I, Ojalvo AG, Seralena A, Pupo E, Lugo V, Pa´ez R. Mutant streptokinase lacking the C-terminal 42 amino acids is less reactive with preexisting antibodies in patient sera. Biochem Biophys Res Commun. 1999;266:230–6.
Torrens I, Ojalvo AG, Seralena A, Hayes O, de la Fuente J. A mutant streptokinase lacking the C-terminal 42 amino acids is less immunogenic. Immunol Lett. 1999;70:213–8.
Fay WP, Bokka LV. Functional analysis of the amino- and carboxyl-termini of streptokinase. Thrombo Haemost. 1998;79:985–91.
Arabi R, Roohvand F, Norouzian D, Sardari S, Aghasadeghi MR, Khanahmad H, et al. A comparative study on the activity and antigenicity of truncated and full-length forms of streptokinase Pol J Microbiol. 2011;60(3):243-51.
Tharp AC, Laha M, Panizzi P, Thompson MW, Fuentes-Prior P, Bock PE. Plasminogen Substrate Recognition by the Streptokinase-Plasminogen Catalytic Complex Is Facilitated by Arg253, Lys256, and Lys257 in the Streptokinase beta domain and Kringle 5 of the Substrate. J Biol Chem. 2009;284(29):19511–21.
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