Cloned (Comment) | Organism |
---|---|
expression of alphaI-tryptase mutants in insect cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
co-crystallization of free and of leupeptin-complexed alphaI-tryptase mutants, 12 mg/ml protein in 10 mM Mops, pH 6.8, 0.7 M NaCl and 0.2 mM leupeptin at room temperature, sitting drop vapor diffusion method, equilibration against reservoir buffer containing 28% PEG 1500, 2 weeks, X-ray diffraction structure determination and anaylsis at 1.6 A resolution, molecular replacement | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D216G | site-directed mutagenesis, alphaI-tryptase mutant, crystal structure determination and analysis, the sole mutation of Asp216 to Gly produces a protease with catalytic and instability properties intermediate between those of alpha and beta-tryptases | Homo sapiens |
K192Q | site-directed mutagenesis, alphaI-tryptase mutant, crystal structure determination and analysis, the single mutation of Lys192 to Gln had no effect on activity or enzyme stability | Homo sapiens |
K192Q/D216G | site-directed mutagenesis, alphaI-tryptase mutant, crystal structure determination and analysis | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
leupeptin | binding structure with alpha- and beta-tryptase, overview | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant alphaI-tryptase mutants from insect cells | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
mast cell | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | beta-tryptase is a tetramer that has enzymatic activity, but requires heparin binding to maintain functional and structural stability, alpha-tryptase has little, if any, enzymatic activity but is a stable tetramer in the absence of heparin, both forms are in a two-state equilibrium, which is influenced by the residues in the vicinity of the active site and by inhibitor/substrate binding, substrate binding structure, overview, Asp216 and Lys192 are the main determinants of tryptase activity | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | beta-tryptase is a tetramer that has enzymatic activity, but requires heparin binding to maintain functional and structural stability, alpha-tryptase has little, if any, enzymatic activity, probably due to a cryptic active site, but is a stable tetramer in the absence of heparin, both forms are in a two-state equilibrium, which is influenced by the residues in the vicinity of the active site and by inhibitor/substrate binding, overview | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
alpha-tryptase | - |
Homo sapiens |
beta-tryptase | - |
Homo sapiens |
tryptase alpha | - |
Homo sapiens |
tryptase beta | - |
Homo sapiens |