Information on EC 3.4.21.64 - peptidase K

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
3.4.21.64
-
RECOMMENDED NAME
GeneOntology No.
peptidase K
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Hydrolysis of keratin, and of other proteins with subtilisin-like specificity. Hydrolyses peptide amides
show the reaction diagram
hydrolysis of keratin, and of other proteins with subtilisin-like specificity
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
EC 3.4.21.14
-
-
formerly
-
EC 3.4.21.14
Tritirachium album Limber
-
formerly
EC 3.4.21.4
-
formerly
endopeptidase K
-
-
mesophilic proteinase K
-
-
Proteinase K
-
-
-
-
Proteinase K
P06875
-
Proteinase K
-
-
Proteinase K
Tritirachium album Limber
-
-
Proteinase, Tritirachium album serine
-
-
-
-
Tritirachium album proteinase K
-
-
-
-
Tritirachium alkaline proteinase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
39450-01-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
commercial preparation from Sigma-Aldrich
-
-
Manually annotated by BRENDA team
Merck strain No. 2429
-
-
Manually annotated by BRENDA team
commercial preparation from Roche Diagnostics
-
-
Manually annotated by BRENDA team
Tritirachium album Limber
-
-
-
Manually annotated by BRENDA team
Tritirachium album Limber
commercial preparation, Amresco
-
-
Manually annotated by BRENDA team
Tritirachium album Limber
commercial preparation, Merck
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
AAPA + H2O
?
show the reaction diagram
-
molecular dynamics simulations of the proteinase K alone and in complex with the peptide substrate AAPA are performed to investigate the effect of substrate binding on the dynamics/molecular motions of proteinase K
-
-
?
Acetyl-Tyr ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Aldolase + H2O
Hydrolyzed aldolase
show the reaction diagram
-
-
-
-
-
Alkynyl carboxylates + H2O
?
show the reaction diagram
-
-
-
-
-
asialofetuin + H2O
?
show the reaction diagram
-
-
-
-
?
Bovine ribonuclease + H2O
Hydrolyzed bovine ribonuclease
show the reaction diagram
-
-
-
-
-
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
hydrolyzed casein
show the reaction diagram
-
-
-
-
-
casein + H2O
?
show the reaction diagram
-
-
-
-
?
creatine kinase + H2O
?
show the reaction diagram
Tritirachium album Limber
-
inactivation of rabbit muscle creatine kinase by processing
-
?
fetuin + H2O
?
show the reaction diagram
-
-
-
-
?
Glucose dehydrogenase + H2O
Hydrolyzed glucose dehydrogenase
show the reaction diagram
-
upon proteolysis the enzyme is inactivated and the polypeptide chain is cleaved into 2 distinct fragments (K-protein, MW 26000 and K-peptide, MW 3000), the cleavage occurs in the C-terminal region of the polypeptide chain.-Leu-Ala-+-Ser-Ser-Glu is proposed as the cleavage site, the term -+- depicts the point of cleavage
upon proteolysis the enzyme is inactivated and the polypeptide chain is cleaved into 2 distinct fragments (K-protein, MW 26000 and K-peptide, MW 3000), the cleavage occurs in the C-terminal region of the polypeptide chain. Leu-Ala-+-Ser-Ser-Glu is proposed as the cleavage site, the term -+- depicts the point of cleavage
-
human growth hormone + H2O
?
show the reaction diagram
-
proteolytic activity and specificity of PK is maintained after its immobilization to magnetic particles
-
-
?
Keratin + H2O
Hydrolyzed keratin
show the reaction diagram
-
-
-
-
-
Keratin + H2O
?
show the reaction diagram
-
-
-
-
?
N-Acetylated amino acid esters + H2O
?
show the reaction diagram
-
-
-
-
-
N-Acetylated peptide esters + H2O
?
show the reaction diagram
-
-
-
-
-
N-succinyl-Ala-Ala-Ala-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Ala + p-nitroaniline
show the reaction diagram
Tritirachium album Limber
-
-
-
?
N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O
N-succinyl-Ala-Ala-Pro-Leu + p-nitroaniline
show the reaction diagram
-
-
-
-
?
Oxidized insulin B-chain + H2O
Hydrolyzed oxidized insulin B-chain
show the reaction diagram
-
main cleavage sites: Gln4-His5, Ser9-His10, Leu11-Val12, Leu15-Tyr16, Leu17-Val18, Phe24-Phe25, Tyr26-Thr27
main cleavage sites: Gln4-His5, Ser9-His10, Leu11-Val12, Leu15-Tyr16, Leu17-Val18, Phe24-Phe25, Tyr26-Thr27
-
p-nitrophenyl acetate + H2O
p-nitrophenol + acetate
show the reaction diagram
Tritirachium album Limber
-
-
-
-
?
poly(L-lactide) + H2O
?
show the reaction diagram
-
enzyme moves on the surface of substrate film to hydrolyze the film around it
-
-
?
prion protein + H2O
?
show the reaction diagram
-
for mouse RML prions, the majority of proteinase K-sensitive disease-related prion protein isoforms do not appear to contribute significantly to infectivity. In human variant Creutzfeldt-Jakob disease, up to 90% of total prion protein present in the brain resists degradation with thermolysin, whereas only 15% of this material resists digestion by proteinase K
-
-
?
pro-recombinant transglutaminase + H2O
?
show the reaction diagram
-
successful cleavage at the pro-sequence
-
-
-
Propynyl benzoate + H2O
?
show the reaction diagram
-
-
-
-
-
ribonuclease A + H2O
proteolytic fragments
show the reaction diagram
Tritirachium album Limber
-
-
-
?
Serum albumin + H2O
Hydrolyzed serum albumin
show the reaction diagram
-
-
-
-
-
succinyl-AAPF-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
Succinyl-Ala-Ala-Ala 2-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
succinyl-Ala-Ala-Ala-p-nitroanilide + H2O
succinyl-Ala-Ala-Ala + p-nitroaniline
show the reaction diagram
-
-
-
-
?
Synthetic peptide substrates + H2O
?
show the reaction diagram
-
primarily specific against aromatic or hydrophobic amino acid residues at the carboxyl side of the splitting point, activity is markedly promoted by elongating the peptide chain to the N-terminal from the splitting point
-
-
-
Urea-denatured hemoglobin + H2O
Hydrolyzed urea-denatured hemoglobin
show the reaction diagram
-
-
-
-
-
Lactate dehydrogenase + H2O
Hydrolyzed lactate dehydrogenase
show the reaction diagram
-
-
-
-
-
additional information
?
-
-
-
-
-
-
additional information
?
-
-
the smallest peptide hydrolyzable should be a tetrapeptide, so that the enzyme could be used as an appropriate tool for sequence analysis of medium size peptides
-
-
-
additional information
?
-
-
the combined action of detergent and proteinase K is effective in degrading `masked' proteins in a poly(adenosine diphosphoribose) preparation which cannot be attacked by the proteinase alone
-
-
-
additional information
?
-
-
specificity for peptide bonds adjacent to the carboxylic group of aliphatic and aromatic amino acids
-
-
-
additional information
?
-
Tritirachium album Limber
-
exhibits a preference for carboxy groups adjacent to aliphatic and aromatic acids and especially those adjacent to alanine residues
-
?
additional information
?
-
-
segment GGG of human prion protein strongly binds as a substrate at the substrate recognition site
-
-
-
additional information
?
-
-
digestion of mouse cell lysates overexpressing prion proteins
-
-
-
additional information
?
-
Tritirachium album Limber
-
PK is very effective in destroying cellular prion proteins and endogenous proteases present in brain homogenate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Keratin + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
segment GGG of human prion protein strongly binds as a substrate at the substrate recognition site
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
the native proteinase K contains two Ca2+ cations
Ca2+
-
proteinase K contains two Ca2+ ions
Calcium
-
2 calcium ions are bound to the native enzyme, activity drops by 70% if this Ca2+ is removed by EDTA; required for folding of the polypeptide chain; the first calcium site is formed by the loop of the residues 174-178 and Asp200; the second more mobile Ca2+ site bridges 2 loops close to the amino and the carboxy termini
Calcium
-
X-ray studies show that it has 2 binding sites for Ca2+, Ca2+ is not directly involved in the catalytic mechanism and is 16.6 A away from the alpha-carbon atoms of the catalytic triad Asp39-His69-Ser224, the activity of the enzyme towards the synthetic substrate succinyl-Ala-Ala-Ala 4-nitroanilide drops slowly to about 20% of its original value when it is depleted of Ca2+
SDS
-
stimulates hydrolysis of serum albumin in a dose-dependent manner, caused primarily by denaturation of the protein substrate, inactivates with an oligopeptide as substrate
Urea
-
stimulates hydrolysis of serum albumin in a dose-dependent manner, caused primarily by denaturation of the protein substrate
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Carbobenzoxy-Ala-Ala-chloromethyl ketone
-
-
Chloromethyl ketone derivatives
-
e.g. carboxybenzoyl-Ala-Gly-PheCH2Cl, carboxybenzoyl-Ala-PheCH2Cl
Cu2+
Tritirachium album Limber
-
at equilibrium two to three copper ions bind stoichiometrically to PK and destroy its activity. Initial reversible and weak binding phase and a slower, irreversible abolition of activity with a half-time of 6 min at saturating copper ion concentrations. PK digestion of cellular prion proteins and other proteins in brain homogenate is inhibited in a concentration-dependent manner at concentrations of more than 1 mM. Presence of calcium ions, up to 10 mM, has no effect on copper inhibition
EDTA
-
22C, or 37C, not inhibitory, 50C, 50% inhibition
EDTA
-
22C, not inhibitory, 37%, 60% residual activity, 50C, complete inhibition
fructose 1,6-diphosphate
-
mixed-type inhibition, more than one inhibitor molecule binds to proteinase K
glucose 6-phosphate
-
mixed-type inhibition, more than one inhibitor molecule binds to proteinase K
Inhibitor from Helix aspersa
-
weak
-
Low-molecular-weight protein proteinase inhibitors from the granule-rich fraction of equine neutrophilic granulocytes
-
-
-
MeOSuc-Ala-Ala-Ala-Pro-Phe-CH2Cl
-
inhibits proteinase K (100 microgram/ml) at concentrations as low as 0.25 mM
MeOSuc-Ala-Ala-Pro-Phe-CH2Cl
-
examination of inhibitory activity using a real-time reverse transcription-polymerase chain reaction assay in the presence of proteinase K. The AAPF inhibitor at a concentration of 0.05 mM allows a signal to be obtained for exogenous target Xeno RNA at 30 cycles
MeOSuc-Ala-Ala-Pro-Val-CH2Cl
-
strong inhibitor
N-Acetyl-L-Pro-L-Ala-L-Pro-L-Phe-D-Ala-L-Ala-NH2
-
substrate analogue
phenylmethylsulfonyl fluoride
-
-
PKI3
-
consists of 180 amino acids with a MW of 19641; natural inhibitor isolated from wheat
-
PKI3
-
crystallization of the inhibitor; natural inhibitor isolated from wheat
-
Polyvalent proteinase inhibitor from albumin gland of Helix pomatia
-
-
-
Proteinase-inhibitors from the albumin gland of Achatina fulica
-
-
-
SDS
-
stimulates hydrolysis of serum albumin in a dose-dependent manner, caused primarily by denaturation of the protein substrate, inactivates with an oligopeptide as substrate
Turkey ovomucoid
-
enzyme interacts with the third domain at the Leu18-Glu19 peptide bond, the reactive site of the inhibitor
-
MeOSuc-Ala-Pro-Ala-Leu-CH2Cl
-
weak inhibitor
additional information
-
not: glucose, fructose
-
additional information
-
not inhibitory: sodium dodecylsulfate
-
additional information
Tritirachium album Limber
-
not inhibited by Zn2+ or Mn2+
-
additional information
-
immobilization of PK on magnetic latex microparticles (500A5 microparticles) and other chemically and structurally similar types of magnetic carriers
-
additional information
-
MeOSuc-Ala-Ala-Ala-Pro-Val-CH2Cl does not inhibit proteinase K (100 microgram/ml) at concentrations as high as 0.75 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
acetonitrile
Tritirachium album Limber
-
-
additional information
Tritirachium album Limber
-
presence of calcium ions, up to 10 mM, has no effect on enzymatic activity
-
additional information
-
highest activity using 3 mg of PK per mg of the carrier
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1
-
acetyl-(Ala)2-Ala methyl ester
-
-
0.8
-
acetyl-(Ala)2-Phe methyl ester
-
-
45
-
Acetyl-Ala methyl ester
-
-
30
-
acetyl-Leu methyl ester
-
-
12
-
acetyl-Phe ethyl ester
-
-
5.6
-
acetyl-Trp ethyl ester
-
-
8.8
-
Acetyl-Tyr ethyl ester
-
-
26
-
acetyl-Val methyl ester
-
-
3.3
-
benzoyl-Arg ethyl ester
-
-
0.9
-
carboxybenzoyl-(Ala)2-Lys methyl ester
-
-
3
-
carboxybenzoyl-Ala-Lys methyl ester
-
carboxybenzoyl-Phe-Lys methyl ester
20
-
carboxybenzoyl-D-Ala-Lys methyl ester
-
-
19
-
carboxybenzoyl-Gly-Lys methyl ester
-
-
9
-
carboxybenzoyl-Leu-Lys methyl ester
-
-
0.46
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 22C
0.48
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 12C
0.52
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 37C
2.36
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 12C
2.46
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 22C
2.76
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 37C
1.29
-
succinyl-Ala-Ala-Ala-p-nitroanilide
-
immobilized PK
4.23
-
succinyl-Ala-Ala-Ala-p-nitroanilide
-
native PK
21
-
carboxybenzoyl-Lys methyl ester
-
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
51
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 12C
88
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 22C
175
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 12C
180
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 37C
364
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 22C
827
-
succinyl-AAPF-4-nitroanilide
-
pH 8.0, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
Tritirachium album Limber
-
specific activity 8.8 U/mg, the activity unit used is absorbance unit, defined as increase of one in absorbance as measured by Lowry's method at 578 nm
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
12
-
urea-denatured hemoglobin
10.5
-
-
-
additional information
-
-
pI: 8.9
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
7
-
proteinase K maintains structural integrity in the range of pH 3.0-7.0
6.7
7.4
Tritirachium album Limber
-
-
8
11
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
70
-
-
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
50
-
soluble and immobilized enzyme
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
from Boehringer, Mannheim
Manually annotated by BRENDA team
-
secretion starts when the stationary phase of growth is reached, and when the culture medium is depleted of glucose and amino acids
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
18500
-
-
Tritirachium album, gel filtration
18500
-
Tritirachium album Limber
-
-
28930
-
-
Tritirachium album, amino acid sequence analysis, the enzyme contains 2 disulfide bonds and a free cysteine residue
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 65500, precursor-protein, calculated, x * 34000, mature protein, SDS-PAGE
monomer
-
1 * 28930, Tritirachium album, amino acid sequence analysis
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
sequence consists of preproregion, catalytic domain and two C-terminal domains. Catalytic domain with the two C-terminal domains is an active protein of 56 kDa that converts at 50C by autolytic cleavage to mature 34 kDa protein
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
at 0.15 nm resolution
-
at 1.5 A resolution
-
at 3.3 A resolution
-
computer modeling studies indicates the presence of a small binding pocket in the vicinity of the proteinase K active site, which may be filled by the aromatic phenylalanine moiety
-
crystallographic study of its complex with a dipeptide chloromethyl ketone inhibitor
-
Langmuir-Blodgett nanotemplate method and hanging drop vapor diffusion method, using 400 mM Na/K-tartrate in 25 mM HEPES pH 7.0
P06875
mercury-inhibited protein in presence of synthetic peptides GGGWGQPH and HGGGW, derived from N-terminal domain of human prion protein. Segment GGG is strongly bound as a substrate at the substrate recognition site
-
structure of the complex of proteinase K with a substrate analogue hexapeptide inhibitor at 2.2 A resolution, N-acetyl-L-Pro-L-Ala-L-Pro-L-Phe-D-Ala-L-Ala-NH2
-
the crystallization of proteinase K using PEG 8000 is performed at various degrees of solvent deuteration and X-ray crystallographic analysis at 1.1 A resolution confirms that deuteration has no effect on crystal quality or crystal structure in this case
-
three-dimensional structure at 1.48 A resolution
-
three-dimensional structure of the complex of proteinase K with its naturally occuring protein inhibitor, PKI3
-
to 1.32 A resolution. final model contains two Ca2+ ions, a molecule of digalacturonic acid and a partially occupied HEPES molecule. The digalacturonic acid molecule has one sugar moiety disposed exactly on a crystallographic twofold axis and is bound through hydrogen-bonding networks involving residue S150 and water molecules
-
to 2.3 A resolution, space group P43212
-
X-ray crystal structure at 1.5 A resolution shows that it has 2 binding sites for Ca2+
-
-
Tritirachium album Limber
-
space group P4(3)2(1)2, cell dimensions a = b = 67.3 A, c = 106.6 A
Tritirachium album Limber
-
space group P4(3)2(1)2, cell dimensions a = b = 68.3, c = 108.4
Tritirachium album Limber
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
-
-
the enzyme acquires partially unfolded conformation at pH 2.5 with lower activity
5.5
9.5
-
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
30 min, about 90% residual activity even in presence of 1% sodium dodecylsulfate
46
-
-
15 min, 50% loss of activity of Ca2+-free enzyme
50
-
-
30 min, 19% residual activity in presence of 1% sodium dodecylsulfate
50
-
-
30 min, 90% residual activity even in presence of 1% sodium dodecylsulfate
60
-
Tritirachium album Limber
-
retains 83% activity after 2 h
65
-
-
15 min, 50% loss of activity of Ca2+-saturated enzyme
66
-
-
thermal unfolding is cooperative at pH 7.0 with transition midpoint of 66C
70
-
-
half-life 30 min
70
-
-
half-life 19 min
additional information
-
-
Ca2+ contributes to the overal stability of the surface regions and improves the thermal stability
additional information
-
-
depletion of Ca2+ increases the rate of autolysis after about 48 h, it reduces the thermal stability
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Depletion of Ca2+ increases the rate of autolysis after about 48 h, it reduces the thermal stability and enhances the deactivation by 8 M urea
-
proteinase K is losing the proteolytic activity as the enzyme is attaining beta conformation
-
Relatively resistant to SDS, 0.2% in 50 mM Tris/HCl, pH 7.4
-
Relatively stable towards heat and denaturing agents
-
Urea, 4 M, stable
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
isopropanol
-
at 40% (v/v) isopropanol nearly half and at 50% (v/v) isopropanol all the tertiary structure is lost, beyond 30% (v/v) isopropanol there is some perturbation in the secondary structure of the protein
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, stable for at least 12 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
mutants purified over a Ni-NTA column
-
-
Tritirachium album Limber
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene encoding proteinase K re-synthesized with an Escherichia coli codon bias and cloned into an arabinose-inducible Escherichia coli expression vector
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A199S
-
no effect on activity
A236V
-
lethal
E138A
-
lethal, increased stability of the serine protease subtilisin
G293A
-
strong positive effect on activity
I132V
-
positive effect on activity
I310K
-
no effect on activity, increased stability of the serine protease subtilisin
K208H
-
positive effect on activity
K332R
-
positive effect on activity
L180I
-
positive effect on activity
L299C
-
lethal, increased stability of the serine protease subtilisin
M145F
-
no effect on activity, increased stability of the serine protease subtilisin
N95C
-
lethal, increased stability of the serine protease subtilisin
P265S
-
no effect on activity, increased stability of the serine protease subtilisin
P355S
-
no effect on activity, increased stability of the serine protease subtilisin
P97S
-
lethal, increased stability of the serine protease subtilisin
R237N
-
no effect on activity
S107D
-
no effect on activity
S123A
-
positive effect on activity
S273T
-
positive effect on activity
S337N
-
positive effect on activity
V167I
-
no effect on activity
V267I
-
positive effect on activity
Y151A
-
strong positive effect on activity
Y194S
-
no effect on activity, random mutation obtained during synthesis of wild-type proteinase K
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
protein engineering approach for increasing activity and heat stability of proteinase K. Protein design algorithms that only require the testing of a small number of variants represent a significant step towards a generic, resource-optimized protein engineering process
biotechnology
-
proteinase K is successfully applied for the activation of purified pro-recombinant transglutaminase either as free or immobilized enzyme and the free enzyme is also applicable directly in the crude cell extract of Escherichia coli. Proteinase K enables a simple two-step activation/purification procedure resulting in protease-free and almost pure transglutaminase preparations
medicine
-
segment GGG of functionally important N-terminal octapeptide region of human prion protein, involved in spongifirm encephalopathy, strongly binds as a substrate at the substrate recognition site
medicine
-
PK magnetic reactor is a useful tool for prion protein digestion
pharmacology
-
In human variant CreutzfeldtJakob disease, up to 90% of total prion protein present in the brain resists degradation with thermolysin, whereas only ?15% of this material resists digestion by proteinase K. Detection of proteinase K-sensitive isoforms of disease-related prion protein using thermolysin should be useful for improving diagnostic sensitivity in human prion diseases
degradation
-
use of enzyme to degrade poly(L-lactide) film. Adsorption of enzyme to film is irreversible, enzyme moves on the surface of substrate to hydrolyze the film around it
medicine
Tritirachium album Limber
-
PK activity in the presence of brain correlates precisely with the concentration of Cu2+ ions that prevent PK digestion of cellular prion protein and other brain proteins. Apparent resistance of cellular prion protein to proteolysis by PK appears to be directly attributable to the inhibition of PK activity by copper-(II) ions