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Information on EC 3.4.21.4 - trypsin and Organism(s) Streptomyces griseus and UniProt Accession P00775
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3.4.21.4 trypsinSpecify your search results
Word Map
- 3.4.21.4
- soybean
- pancreas
- elastase
- pepsin
- thrombin
- amylase
- proteinases
- albumin
- lipase
- porcine
- collagenase
- pronase
- erythrocyte
- plasmin
- collagen
- kallikreins
- sephadex
- neuraminidase
- bean
- lectin
-
exocrine
- carboxypeptidase
- juice
- cathepsins
- coagulation
-
edman
- casein
- duodenal
- plasminogen
-
acinar
- aprotinin
-
cyanogen
- subtilisin
- zymogen
- heparin
-
radioimmunoassay
- midguts
- cholecystokinin
-
reversed-phase
- lysozyme
- venom
-
125i-labeled
- hydrolysates
-
protease-activated
- kinin
- tryptase
- flour
- phosphopeptides
-
hemagglutination
- hyaluronidase
- biotechnology
- synthesis
- food industry
- medicine
- analysis
The taxonomic range for the selected organisms is: Streptomyces griseus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
trypsin, at-ii, cationic trypsinogen, beta-trypsin, trypsin-like enzyme, mesotrypsin, trypsin a, cationic trypsin, anionic trypsinogen, anionic trypsin, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
alpha-trypsin
-
-
-
-
Anionic trypsinogen
-
-
-
-
beta-trypsin
-
-
-
-
Brain trypsinogen
-
-
-
-
Cationic trypsinogen
-
-
-
-
cocoonase
-
-
-
-
Mesotrypsinogen
-
-
-
-
parenzyme
-
-
-
-
parenzymol
-
-
-
-
pseudotrypsin
-
-
-
-
SET
-
-
-
-
sperm receptor hydrolase
-
-
-
-
tripcellim
-
-
-
-
tryptar
-
-
-
-
tryptase
-
-
-
-
trypure
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
CAS REGISTRY NUMBER
COMMENTARY
9002-07-7
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
N-benzoyl-DL-arginine-4-nitroanilide + H2O
N-benzoyl-DL-arginine + 4-nitroaniline
-
-
-
?
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide + H2O
benzyloxycarbonylglycyl-L-prolyl-L-arginine + 7-amino-4-methylcoumarin
-
-
-
-
?
additional information
?
-
-
three-dimensional structure and topological specificities, substrate recognition, overview. The neighboring residues Leu71 and Gln72 in the N-terminal beta-barrel domain of the enzyme synergistically play an important role in substrate recognition. Identification of amino acid residue(s) related to collagen binding for wild-type and mutant enzymes
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Na+
-
Na+ activation is interwoven with substrate selectivity in the trypsin scaffold
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
p-tosyl-L-Lys-chloromethane
-
reaction towards B chain of insulin is inhibited, the activity towards the A-chain is not significantly affected
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Na+
wild-type, no activation, engineering of wild-type into a Na+-activated enzyme by replacing residues in the 170, 186, and 220 loops to those of coagulation factor Xa
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0222
factor Xa
mutant G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C
-
0.285
factor Xa
mutant Y172S/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C
-
0.002
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, recombinant enzyme
0.0023
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, wild-type enzyme
0.0032
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, recombinant enzyme
0.0036
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, wild-type enzyme
0.0044
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190A
0.0061
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190S
0.0101
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190S
0.0129
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190A
0.067
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190P
0.166
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190P
0.224
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190V
0.231
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190V
0.012
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y/Q72R, pH 8.0, 37°C
0.0172
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y, pH 8.0, 37°C
0.0177
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
wild-type enzyme, pH 8.0, 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
9.25
factor Xa
mutant G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C
-
77.7
factor Xa
mutant Y172S/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C
-
0.52 - 0.7
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, wild-type enzyme
2.7
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, recombinant enzyme
8
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190P
38.33
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190V
82.5
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190A
0.04 - 1.97
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190P
0.5
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190V
3.2
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190A
8.78
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
pH 7.6, mutant enzyme T190P
33
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
wild-type enzyme, pH 8.0, 37°C
51.3
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y, pH 8.0, 37°C
57.3
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y/Q72R, pH 8.0, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0019
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
wild-type enzyme, pH 8.0, 37°C
0.0032
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y, pH 8.0, 37°C
0.0048
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
-
mutant L71Y/Q72R, pH 8.0, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.34
after 1fold purification, using N-benzoyl-DL-arginine-4-nitroanilide as a substrate
9.42
after 27.6fold purification, using N-benzoyl-DL-arginine-4-nitroanilide as a substrate
110.2
-
mutant L71Y/Q72R, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
66.1
-
wild-type enzyme, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
90.3
-
mutant L71Y, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
23080
mutant enzyme T190A, electrospray mass spectrometry analysis
23090
mutant enzyme T190S, electrospray mass spectrometry analysis
23100
23110
23100
mutant enzyme T190P, electrospray mass spectrometry analysis
23100
mutant enzyme T190V, electrospray mass spectrometry analysis
23110
wild-type enzyme and recombinant enzyme, electrospray mass spectrometry analysis
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
three-dimensional structure and topological specificities, substrate recognition, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, recombinant wild-type enzyme and mutant enzyme T190P at 1.5 and 1.9 A resolution
FX99 mutant of trypsin, hanging drop vapour diffusion method, using 1.6 M ammonium sulfate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A171S/Y172S/G173S/N174F/E175I/E180M/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
contrary to wild-type, mutant may be activated by Na+, mutations in 170 loop increase susceptibility to Na+ further
G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
twofold increase in KM-value
T190A
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 47% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 55% of that of the wild-type enzyme
T190P
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 0.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.7% of that of the wild-type enzyme
T190S
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 18% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 9% of that of the wild-type enzyme
T190V
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 2.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.6% of that of the wild-type enzyme
Y172M/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
contrary to wild-type, mutant may be activated by Na+
Y172S/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
contrary to wild-type, mutant may be activated by Na+
Y172S/Y217E/P222K/Y224K/P225Y
30fold increase in KM-value, contrary to wild-type, mutant may be activated by Na+
L71Y
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type
L71Y/Q72R
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type
Q72R
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The mutant shows slightly lower activities toward type I collagen than the wild-type
additional information
-
the FX99 mutant is constructed using 19 replacements in wild type trypsin with residues present in FXa; i.e. N95T/G96K/T97E/insertionT98/insertionY99 in the 99-loop, A171S/Y172S/G173S/N174F/E175I/E180M in the 170-loop, DELTAP185/G186K/G187Q/V188E in the 186-loop, and Y217E/P222K/Y224K/P225Y in the 220-loop
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10
-
20% loss of activity after 12 h at 0°C, complete loss of activity after 25 min at 30°C
95441
additional information
-
significant stabilization in the neutral and alkaline range by 15 mM Ca2+
95441
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
30
-
pH 8.0, 75% loss of activity after 25 min, 90% loss of activity after 24 h
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
significant stabilization in the neutral and alkaline range by 15 mM Ca2+
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
SP-Sepharose Fast Flow column chromatography and Arg-Sepharose 4B Fast Flow column chromatography
recombinant enzyme from Streptomyces lividans strain 1326 culture supernatant by cation exchange chromatography and dialysis
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme expression from expression vector pTONA5a, which includes a promoter from Streptomyces metalloendopeptidase, in Streptomyces lividans strain 1326
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Read, R.J.; James, M.N.G.
Refined crystal structure of Streptomyces griseus trypsin at 1.7 A resolution
J. Mol. Biol.
200
523-551
1988
Streptomyces griseus
Olafson, R.W.; Smilie, L.B.
Enzymic and physicochemical properties of Streptomyces griseus trypsin
Biochemistry
14
1161-1167
1975
Streptomyces griseus
Page, M.J.; Wong, S.L.; Hewitt, J.; Strynadka, N.C.; MacGillivray, R.T.
Engineering the primary substrate specificity of Streptomyces griseus trypsin
Biochemistry
42
9060-9066
2003
Streptomyces griseus (P00775), Streptomyces griseus
Page, M.J.; Bleackley, M.R.; Wong, S.; MacGillivray, R.T.; Di Cera, E.
Conversion of trypsin into a Na(+)-activated enzyme
Biochemistry
45
2987-2993
2006
Streptomyces griseus (P00775), Streptomyces griseus
Stosova, T.; Sebela, M.; Rehulka, P.; Sedo, O.; Havlis, J.; Zdrahal, Z.
Evaluation of the possible proteomic application of trypsin from Streptomyces griseus
Anal. Biochem.
376
94-102
2008
Streptomyces griseus (P00775), Streptomyces griseus
Page, M.J.; Carrell, C.J.; Di Cera, E.
Engineering protein allostery: 1.05 A resolution structure and enzymatic properties of a Na+-activated trypsin
J. Mol. Biol.
378
666-672
2008
Streptomyces griseus
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