Studies on Lipidification of Streptokinase

Abstract

Thrombotic disorders and their associated problems are extensively prevalent in developed and developing countries. Streptokinase (SK) is a well-known thrombolytic agent, which is very useful in treating coronary thrombosis and acute myocardial infarction. Several attempts have been made to date to make improvements of this wonderful molecule in terms of reducing or eliminating the problems of eliciting immunogenicity and enhancing the half-life of the molecule. The present research is focused to produce a recombinant SK with enhanced stability and biological activity by the methodology of lipid modification. SK was targeted successfully to the membrane with the help of modified apyrase signal sequence. Higher expression was reported for GJ1158 strain in LBON medium when compared with BL21 (DE3). The obtained recombinant SK was tested for its biological activity by the method of caseinolytic assay. The higher clearance zone was observed in recombinant lipid-modified streptokinase, which denotes the enhanced activity of the protein. The present trial of lipid modification of therapeutics, particularly SK, could help for its superior use as a thrombolytic agent and also paves way for many of the other clinical applications.

Incorporation of a tag helps to overcome expression variability in a recombinant host

“…Solubility of proteins is proposed to be inversely correlated with accumulation due to increased accessibility to host proteases [13] , [20] , [23] , [27] . The solubility profile of histones expressed using pET28a vector in BL21 (DE3) pLysS strain at 37 °C is depicted in Fig.…”

Structural Biology and Protein Engineering of Thrombolytics

“…Preventing the cleavage by mutagenesis at the identified positions (Lys59Gln/Glu, Lys386Gln) resulted in a 21-fold increased half-life without affecting activity [[235], [236], [237]]. Alternative strategies based on combinatorial mutagenesis [14,238], PEGylation [[239], [240], [241], [242]], glycosylation [235,243], or lipidification [244] yielded streptokinase variants with improved thrombolytic activity, decreased immunogenicity, and higher half-life.…”