Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Homo sapiens |
into the BamH/HindIII restriction sites of the pQE30 vector and expressed in Escherichia coli M15[pREP4] cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
by vapor diffusion in hanging drops. K4 crystallised in the presence of 10 mM Ni2+ reveals tetragonal space group P4. In the presence of Zn2+ it reveals tetragonal space group P41212. In the presence of 10 mM Co2+ it reveals monoclinic space group P21. The three different K4 crystal forms with bound Ni2+, Zn2, and Co2+ contain two, four and 16 crystallographically independent K4 molecules per asymmetric unit, respectively. K4 possesses an unusual 99-loop that creates a groove-like acidic S2 subsite | Homo sapiens |
hanging drop vapour diffusion method using 20% PEG-MME 2000, 100 mM Tris (pH 8.5) and 10 mM NiCl2, or 100 mM HEPES (pH 6.5) and 100 mM NaCl with 1.6 M ammonium sulfate, or with 10 mM CoCl2, 100 mM Mes buffer (pH 6.0) and 1.6 M ammonium sulfate | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E77A | the mutation completely abolishes the inhibitory effect of Zn2+ for hK4 | Homo sapiens |
E77A | zinc-independent | Homo sapiens |
H25A | the mutation completely abolishes the inhibitory effect of Zn2+ for hK4 | Homo sapiens |
H25A | zinc-independent | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Co2+ | - |
Homo sapiens | |
Ni2+ | - |
Homo sapiens | |
Zn2+ | enzyme activity is high at zinc concentrations below 0.001 mM and decreases to about 30% at 0.1 mM. Zinc affects the K4 active site via the salt-bridge formed between the N terminus and Asp194 required for a functional active site | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0207 | - |
Z-FVR-p-nitroanilide | in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25°C | Homo sapiens | |
0.0207 | - |
Z-Phe-Val-Arg-p-nitroanilide | monomeric K4 | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23997 | - |
1 * 25000, SDS-PAGE, 1 * 23997, MALDI-TOF mass spectrometry | Homo sapiens |
24000 | - |
MALDI-TOF mass spectrometry | Homo sapiens |
25000 | - |
SDS-PAGE | Homo sapiens |
700000 | - |
gel filtration, aggregate of four octamers | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
by Ni-NTA chromatography | Homo sapiens |
nickel-nitrilotriacetic acid-Sepharose column chromatography | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
ovary cancer cell | - |
Homo sapiens | - |
prostate cancer cell | - |
Homo sapiens | - |
prostate gland | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Tos-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O | - |
Homo sapiens | ? | - |
? | |
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O | - |
Homo sapiens | tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin | - |
? | |
Z-FVR-p-nitroanilide + H2O | - |
Homo sapiens | Z-FVR + p-nitroaniline | - |
? | |
Z-Phe-Val-Arg-p-nitroanilide + H2O | - |
Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 25000, SDS-PAGE, 1 * 23997, MALDI-TOF mass spectrometry | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
HK4 | - |
Homo sapiens |
K4 | - |
Homo sapiens |
kallikrein 4 | - |
Homo sapiens |
KLK-L1 | - |
Homo sapiens |
KLK4 | - |
Homo sapiens |
peptidase S01.251 | - |
Homo sapiens |
PRSS17 | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.64 | - |
Z-FVR-p-nitroanilide | in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25°C | Homo sapiens | |
3.64 | - |
Z-Phe-Val-Arg-p-nitroanilide | monomeric K4 | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.016 | - |
in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25°C | Homo sapiens | Zn2+ | |
0.0164 | - |
permanent residual activity | Homo sapiens | Zn2+ | |
0.1 | - |
- |
Homo sapiens | Ni2+ | |
0.1 | - |
in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25°C | Homo sapiens | Ni2+ |