Literature summary for 3.4.21.B12 extracted from

Debela, M.; Magdolen, V.; Grimminger, V.; Sommerhoff, C.; Messerschmidt, A.; Huber, R.; Friedrich, R.; Bode, W.; Goettig, P.
Crystal structures of human tissue kallikrein 4: activity modulation by a specific zinc binding site (2006), J. Mol. Biol., 362, 1094-1107.

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Cloned(Commentary)

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 (Commentary)

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

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

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

Inhibitors	Comment	Organism
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 [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 [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

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
by Ni-NTA chromatography	Homo sapiens
nickel-nitrilotriacetic acid-Sepharose column chromatography	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
ovary cancer cell	-	Homo sapiens	-
prostate cancer cell	-	Homo sapiens	-
prostate gland	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Subunits	Comment	Organism
monomer	1 * 25000, SDS-PAGE, 1 * 23997, MALDI-TOF mass spectrometry	Homo sapiens

Synonyms

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 [1/s]

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	IC50 Value Maximum	Comment	Organism	Inhibitor
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+