Streptokinase—a clinically useful thrombolytic agent

doi:10.1016/j.biotechadv.2003.09.004

Biotechnology Advances

Volume 22, Issue 4, February 2004, Pages 287-307

https://doi.org/10.1016/j.biotechadv.2003.09.004 Get rights and content

Abstract

A failure of hemostasis and consequent formation of blood clots in the circulatory system can produce severe outcomes such as stroke and myocardial infraction. Pathological development of blood clots requires clinical intervention with fibrinolytic agents such as urokinase, tissue plasminogen activator and streptokinase. This review deals with streptokinase as a clinically important and cost-effective plasminogen activator. The aspects discussed include: the mode of action; the structure and structure–function relationships; the structural modifications for improving functionality; recombinant streptokinase; microbial production; and recovery of this protein from crude broths.

Introduction

A blood clot (thrombus) developed in the circulatory system can cause vascular blockage leading to serious consequences including death. A healthy hemostatic system suppresses the development of blood clots in normal circulation, but reacts extensively in the event of vascular injury to prevent blood loss. Outcomes of a failed hemostasis include stroke, pulmonary embolism, deep vain thrombosis and acute myocardial infraction. Pathologies involving a failure of hemostasis and the development of clot require clinical intervention consisting of intravenous administration of thrombolytic agents Collen et al., 1988, Collen, 1990, Francis and Marder, 1991. Streptokinase is one such agent. Other thrombolytic or fibrinolytic agents include urokinase and the tissue type plasminogen activator (tPA). This review outlines the process of clot dissolution (i.e., thrombolysis or fibrinolysis) and the various thrombolytic agents available to achieve it. One clinically important thrombolytic agent, streptokinase, and its production from microorganisms are discussed in detail.

Numerous trails have been conducted to compare the clinical efficacy of recombinant tPA and streptokinase. Generally, these investigations have not revealed a clear preference for either drug. Streptokinase is as effective as recombinant tPA in treating acute myocardial infarction (Sherry and Marder, 1991), and it is certainly more cost-effective; however, its use is not risk free. In view of the relatively recent availability of the competing recombinant tPA, skepticism is being expressed about the continued viability of streptokinase therapy Sane and Little, 1998, Werf, 1999. Despite this, research on streptokinase continues, and it remains a vital affordable therapy specially in the world's poorer healthcare systems.

Section snippets

Thrombolysis or fibrinolysis

The physiology of the fibrin–clot formation is relatively well understood Paoletti and Sherry, 1977, Wu and Thiagarajan, 1996. A blood clot or thrombus consists of blood cells occluded in a matrix of the protein fibrin. Enzyme-mediated dissolution of the fibrin clot is known as thrombolysis or fibrinolysis. In mammalian circulation, the enzyme responsible for fibrinolysis is plasmin, a trypsin-like serine protease Castellino, 1981, Rouf et al., 1996. The fibrinolytically active plasmin is

Comparing plasminogen activators

Both tPA and uPA are trypsin-like serine proteases which activate plasminogen directly. They have the respective molecular weights of 70 and 55 kDa. Both tPA and uPA are glycoproteins. uPA is produced by the kidneys and secreted into the urine Barlow, 1976, Paoletti and Sherry, 1977, whereas tPA is produced by the vascular endothelial cells Camiolo et al., 1971, Strassburger et al., 1983.

tPA for therapeutic use is obtained mainly from cultures of recombinant animal cells (Rouf et al., 1996).

Streptokinase

The extracellular enzyme streptokinase (EC 3.4.99.22) is produced by various strains of β-hemolytic streptococci. The enzyme is a single-chain polypeptide that exerts its fibrinolytic action indirectly by activating the circulatory plasminogen. The complete amino acid sequence of streptokinase was first established by Jackson and Tang (1982). Streptokinase has a molar mass of 47 kDa and is made up of 414 amino acid residues (Malke and Ferretti, 1984). The protein exhibits its maximum activity

Producing microorganisms

Streptokinase producing streptococci were first identified in 1874 by Billroth in exudates of infected wounds. Later, the blood of scarlet fever patients was shown to contains similar microorganisms. By 1919, Streptococcus sp. had been classified into alpha, beta and gamma variants based on the distinct types of hemolytic reactions the variants produced on blood agar plates.

In 1933, Lancefield used serologic distinctions to further differentiate the β-hemolytic streptococci into groups A to O

Concluding remarks

Native streptokinase is useful for cost-effective thrombolytic therapy in clinical practice, but its use is not risk free. Large quantities of streptokinase can be produced inexpensively via bacterial fermentation. Cloning of the streptokinase gene in nonpathogenic microorganisms has enabled production of recombinant streptokinase that eliminates any risk of inadvertent inoculation of patients and production personnel with potentially pathogenic streptococci.

Various chemical modifications of

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Research review paperStreptokinase—a clinically useful thrombolytic agent

Abstract

Introduction

Section snippets

Thrombolysis or fibrinolysis

Comparing plasminogen activators

Streptokinase

Producing microorganisms

Concluding remarks

J. Biol. Chem.

J. Biol. Chem.

Methods Enzymol.

J. Biol. Chem.

Methods Enzymol.

Fibrinolysis

Biochim. Biophys. Acta

J. Biol. Chem.

Biochim. Biophys. Acta

Prog. Cardiovasc. Dis.

Enzyme Microb. Technol.

Anal. Biochem.

Int. J. Cardiol.

Thromb. Res.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

FEBS Lett.

Med. Anal. Biochem.

J. Biol. Chem.

Circulation

Zentralbl. Bakteriol.

Gene

Methods

J. Biol. Chem.

Blood

Int. J. Biol. Macromol.

FEMS Microbiol. Lett.

J. Biol. Chem.

J. Am. Coll. Cardiol.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Affinity chromatographic purification of streptokinase with monoclonal antibodies

Allerg. Immunol. (Leipzig)

Cloning, expression and purification of recombinant streptokinase: partial characterization of the protein expressed in Escherichia coli

Braz. J. Med. Biol. Res.

Expression and characterization of the intact N-terminal domain of streptokinase

Protein Sci.

Unfolding and aggregation during the thermal denaturation of streptokinase

Eur. J. Biochem.

Microbiological isolation of a protease with fibrinolytic effect from Aspergillus ochraceus

Folia Haematol.

Multidomain structure of a recombinant streptokinase. A differential scanning calorimetry study

J. Protein Chem.

An improved medium for the cultivation of hemolytic streptococcus

J. Bacteriol.

Clinical relevance of anti-thrombin-III

Semin. Thromb. Hemost.

Streptokinase binds preferentially to the extended conformation of plasminogen through lysine binding site and catalytic domain interactions

Biochemistry (U.S.)

A characterization of native streptokinase and altered streptokinase isolated from a human plasminogen activator complex

Biochemistry

Fibrinogenolysis and fibrinolysis with tissue plasminogen activator, urokinase, streptokinase-activated human globulin, and plasmin

Proc. Soc. Exp. Biol. Med.

Recent advances in the chemistry of the fibrinolytic system

Chem. Rev.

Biochemistry of human-plasminogen

Semin. Thromb. Hemost.

Strategies in downstream processing

Biosafety

Streptococcal fibrinolysis: a proteolytic reaction due to serum enzyme activated by streptococcal fibrinolysin

J. Gen. Physiol.

Methods for measuring the activity of components of the Streptococcal fibrinolytic system and Streptococcal desoxyribonuclease

J. Clin. Invest.

Research review paper
Streptokinase—a clinically useful thrombolytic agent