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Literature summary for 3.4.21.22 extracted from

  • Kristensen, L.H.; Olsen, O.H.; Blouse, G.E.; Brandstetter, H.
    Releasing the brakes in coagulation factor IXa by co-operative maturation of the substrate-binding site (2016), Biochem. J., 473, 2395-2411 .
    View publication on PubMed
Search for more literature for 3.4.21.22

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli NiCo21(DE3) cells Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
activated mutant K98T, sitting drop vapor diffusion method, using 0.1 M MES (pH 6.5) and 18-24% (w/v) PEG6000 Homo sapiens

Protein Variants

Protein Variants Comment Organism
K98T the variant demonstrates a 2fold improvement in kcat/KM value for Pefa9 as compared to the wild type enzyme Homo sapiens
K98T/R150A/I213V the mutations result in a 4fold increase in kinetic efficiency compared with the wild type enzyme Homo sapiens
R150A the mutation reduces potential autolysis in the 148-loop during production and leads to reduced activity compared to the wild type enzyme Homo sapiens
R150A/I213V the variant shows increases in the catalytic efficiency for amidolytic activity when the target substrate is either Pefa9 or Pefa10 with 1.5fold and 1.2fold increases, respectively Homo sapiens
V16I/I21/V R150A the variant results in a negligible increase in activity compared with the V16I/R150A variant Homo sapiens
V16I/K98T/I213V the mutations result in a 4fold increase in kinetic efficiency compared with the wild type enzyme Homo sapiens
V16I/K98T/I213V/R150A the mutations result in a 6fold increase in kinetic efficiency compared with the wild type enzyme Homo sapiens
V16I/K98T/R150A the mutant shows enhancements in activity leading to a 2.5fold increased kinetic efficiency compared with the wild type enzyme Homo sapiens
V16I/K98T/Y177T/I213V the variant reduces the kinetic efficiency to the 4fold increase that is also observed with the K98T/R150A/I213V variant Homo sapiens
V16I/R150A the variant shows no increase in the kcat/KM value for Pefa9 as compared to the wild type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
1,5-dansyl-Glu-Gly-Arg chloromethylketone
-
 Homo sapiens
4-aminobenzamidine
-
 Homo sapiens
D-Phe-Pro-Arg chloromethylketone
-
 Homo sapiens
Glu-Gly-Arg chloromethylketone
-
 Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens P00740
-
-

Purification (Commentary)

Purification (Comment) Organism
HiPrep Q column chromatography and Superdex 75 gel filtration Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(methylsulfonyl)-D-cyclohexylalanine-Gly-Arg-7-amido-4-methylcoumarin + H2O Pefa10 Homo sapiens (methylsulfonyl)-D-cyclohexylalanine-Gly-Arg + 7-amino-4-methylcoumarin
-
 ?
(methylsulfonyl)-D-cyclohexylglycine-Gly-Arg-7-amido-4-methylcoumarin + H2O Pefa9 Homo sapiens (methylsulfonyl)-D-cyclohexylglycine-Gly-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butoxycarbonyl-Ile-Glu-Gly-Arg-7-amido-4-methylcoumarin + H2O
-
 Homo sapiens tert-butoxycarbonyl-Ile-Glu-Gly-Arg + 7-amino-4-methylcoumarin
-
 ?

Synonyms

Synonyms Comment Organism
activated factor IX
-
 Homo sapiens
factor XIa
-
 Homo sapiens