Cloned (Comment) | Organism |
---|---|
recombinant expression of the isolated serine protease domain (residues 276-527) wild-type and mutant S478A/C395A/N448Q of enzyme tPA in Pichia pastoris strain X-33. The Pichia pastoris Kex2p protease can cleave the secreted tPA-serine protease domain | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified enzyme in complex with plasminogen activator inhibitor-1, X-ray diffraction structure determination and analysis | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
additional information | engineering on tPA to reduce its inhibition by PAI-1 without compromising its thrombolytic effect is a continuous effort | Homo sapiens |
S478A/C395A/N448Q | site-directed mutagenesis in the serine protease domain | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
plasminogen activator inhibitor-1 | PAI-1, endogenous PAI-1 inactivates the enzyme. Physiologically, PAI-1 inhibits plasminogen activators rapidly and irreversibly, PAI-1 inhibitory mechanism. Analysis of the overall structure of tPA-PAI-1 Michaelis complex, structure-function relationship, overview. The PAI-1 reactive center loop adopts a unique kinked conformation, and on the tPA side, the S2 and S1beta pockets open up to accommodate PAI-1. The deterined crystal structure provides detailed interactions between tPA 37- and 60-loops with PAI-1, overview Comparison to urokinase-type plasminogen activator (uPA, EC 3.4.21.73) structure with bound plasminogen activator inhibitor-1. As a result of the complex formation, an interface of 1202 A2 (solvent-inaccessible area) between PAI-1 and tPA-SPD is formed, which is higher than that of the uPA-SPDx02PAI-1 complex (1058 A2). Of the total area of PAI-1, 8% is involved in interacting with tPA, whereas for uPA, only 6.9% is at the interface. As a protease inhibitor, PAI-1 is very specific to tPA and uPA. Analysis of PAI-1 mutant 14-1B (N150H/K154T/Q319L/M354I) | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P00750 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | Pichia pastoris Kex2p protease can cleave the secreted tPA-serine protease domain | Homo sapiens |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant isolated serine protease domain (residues 276-527)of enzyme tPA from Pichia pastoris strain X-33 by cation exchange chromatography and gel filtration | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
PLAT | - |
Homo sapiens |
Tissue-type plasminogen activator | - |
Homo sapiens |
tPA | - |
Homo sapiens |