The pharmacological experiments on isolated organs (ex vivo) are the preferred method for assessing primary pharmacodynamics of studied drugs, since this is completely excluded systemic influence neurohumoral regulation. In last decade, a new group thrombolytic drugs based immobilized subtilisins has been formed. At stage registrational preclinical and clinical studies, their pleiotropic effect...

Through three cumulative amino acid substitutions, we constructed novel mutant subtilisins E of Bacillus subtilis, all with high specificity, activity and productivity. The substitution of conserved Gly127, constituting P1 substrate-binding pocket, with Ala and Val showed a marked preference for the small P1 substrate. Leu was then substituted for Ile31 next to the catalytic Asp32 to enhance th...

Bacillus subtilisin has been a popular model protein for engineering altered substrate specificity. Although some studies have succeeded in increasing the specificity of subtilisin, they also demonstrate that high specificity is difficult to achieve solely by engineering selective substrate binding. In this paper, we analyze the structure and transient state kinetic behavior of Sbt160, a subtil...

The alkaline proteinase from the mesophilic bacterium Bacillus mesentericus has been crystallized in a 1:1 complex with the inhibitor eglin-C from the medical leech. The crystals have cell dimensions of a = 43.0, b = 71.9, c = 48.3 A and beta = 110.0 degrees and are in the space group P2(1). Three-dimensional data to 2.0 A have been recorded on film from a single crystal. The orientation and po...

Previous work on the primary structures of subtilisins BPN’ (1, 2), Novo (3), and Carlsberg (4) had shown that tryptic digestion and resolution of the resulting peptides by either ion exchange chromatography or gel filtration served as a useful first step in determining the amino acid sequences of these proteins. Accordingly, the diisopropylphosphoryl derivative of subtilisin Amylosacchariticus...

Proteolytic processing plays a significant role in the process of invasion by the obligate intracellular parasite Toxoplasma gondii. We have cloned a gene, TgSUB1, encoding for a subtilisin-type serine protease found in T. gondii tachyzoites. TgSUB1 protein is homologous to other Apicomplexan and bacterial subtilisins and is processed within the secretory pathway of the parasite. Initial cleava...

The activities of Carlsberg and Novo subtilisins towards a number of N-acetylamino acid esters and amino acid esters have been examined. The enzymes were also compared with respect to their efficiency in catalyzing aminblysis reactions, their rates of inactivation by certain aromatic sulfonyl halides, and the rates of deacylation of their N-frans-cinnamoyl derivatives. The enzymes were found to...