Information on EC 3.4.14.9 - tripeptidyl-peptidase I

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
3.4.14.9
-
RECOMMENDED NAME
GeneOntology No.
tripeptidyl-peptidase I
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Release of an N-terminal tripeptide from a polypeptide, but also has endopeptidase activity.
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
aminopeptidase, tripeptidyl, I
-
-
-
-
CLN2
-
-
CLN2 protein
-
-
LPIC
-
-
-
-
lysosomal pepstatin insensitive protease
-
-
-
-
SedB
Q70J58, Q70J59
-
sedolisin B
Q70J59
-
sedolisin C
Q70GH4
-
sedolisin D
Q70J58
-
TPP I
-
prefers Leu, Phe and Nle at the P1 position, whereas Asn, His, Lys, Arg, Ser, Val, Ile, Thr, Gly and Pro are highly unfavored in this position, showing less than 1% of the activity of the best substrates
TPP-I
-
-
Tpp1
O14773
-
Tpp1
O14773
TPP1 is a serine protease that possesses two catalytic functions, a primary tripeptidyl exopeptidase activity with a pH optimum of about 5.0 that catalyzes the sequential release of tripeptides from the unsubstituted N termini of substrates and a much weaker endoproteolytic activity with a pH optimum of about 3.0
TPPI
O14773
-
tripeptidyl aminopeptidase
-
-
-
-
tripeptidyl aminopeptidase I
-
-
-
-
tripeptidyl peptidase
-
-
-
-
tripeptidyl peptidase 1
-
-
tripeptidyl peptidase 1
O14773
-
tripeptidyl peptidase 1
-
-
tripeptidyl peptidase I
-
-
-
-
tripeptidyl peptidase I
-
-
tripeptidyl peptidase I
O14773
-
tripeptidyl peptidase I
-
-
tripeptidyl peptidase I
-
-
tripeptidyl peptidase I
-
shows tripeptidyl peptidase activity and pepstatin insensitive carboxyl endopeptidase activity
tripeptidyl peptidase-I
-
-
tripeptidyl-peptidase 1
-
-
tripeptidyl-peptidase 1
O14773
-
tripeptidyl-peptidase I
-
-
CAS REGISTRY NUMBER
COMMENTARY
151662-36-1
-
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
TPP1 knockout mice which serve as a mouse model for classical late-infantile neuronal ceroid lipofuscinosis (LINCL) are analysed in terms of storage material present in the brain of the mouse model. It is shown that a number of protein constituents including glial fibrillary acidic protein are elevated
malfunction
-
TPP1 knockout mice providing a mouse model for late-infantile neuronal ceroid lipofuscinosis (LINCL) generated that either lack the pro-apoptotic p53 or have increased levels of anti-apoptotic Bcl-2. Neither modification affects the shortened life-span of the LINCL mouse. These findings suggest that targeting pathways of cell death involving p53 or Bcl-2 do not represent useful directions for developing effective treatment
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(pGlu)LYENKPRRRPYIL + H2O
(pGlu)L + YENKPRRRPYIL
show the reaction diagram
-
i.e. neurotensin, 15.0% degradation after 16 h
-
?
Ala-Ala-Ala-4-nitroanilide + H2O
Ala-Ala-Ala + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Ala-Ala-4-nitroanilide + H2O
Ala-Ala-Ala + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-4-nitroanilide + H2O
Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-4-nitroanilide + H2O
Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-4-nitroanilide + H2O
Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedB
-
-
?
Ala-Ala-Phe-4-nitroanilide + H2O
Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedC
-
-
?
Ala-Ala-Phe-4-nitroanilide + H2O
Ala-Ala-Phe + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedD
-
-
?
Ala-Ala-Phe-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-carbamoylmethylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
33.8% degradation within 60 min
-
-
?
Ala-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Ala-Ala-Pro-4-nitroanilide + H2O
Ala-Ala-Pro + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Arg-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Ala-Asp-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Asp-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Ala-His-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-His-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Ala-Nle-Leu-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nle-Leu + 7-amino-4-carbamoylmethylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Nle-Nle-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nle-Nle + 7-amino-4-carbamoylmethylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Nva-Nle-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nva-Nle + 7-amino-4-carbamoylmethylcoumarin
show the reaction diagram
-
-
-
-
?
Ala-Phe-Pro-4-nitroanilide + H2O
Ala-Phe-Pro + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Ala-Pro-Gly-Asp-Arg-Ile-Tyr-Val-His-Pro-Phe + H2O
Ala-Pro-Gly + Asp-Arg-Ile + Tyr-Val-His-Pro-Phe
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedB
-
-
?
Ala-Ser-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Ser-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
angiotensin II + H2O
? + Asp-Arg-Val
show the reaction diagram
-
-
-
-
?
Arg-Ala-Phe-7-amido-4-methylcoumarin + H2O
Arg-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Arg-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Arg-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Arg-Pro-Phe-7-amido-4-carbamoylmethylcoumarin + H2O
Arg-Pro-Phe + 7-amino-4-carbamoylmethylcoumarin
show the reaction diagram
-
-
-
-
?
Asp-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Asp-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
Cholecystokinin + H2O
?
show the reaction diagram
-
-
-
-
?
DRVYIHPF + H2O
DRV + YIHPF
show the reaction diagram
-
-
-
?
DRVYIHPF + H2O
DRV + YIHPF
show the reaction diagram
-
i.e. angiotensin II, 18.2% degradation after 16 h
-
?
DRVYIHPFHL + H2O
DRV + YIHPFHL
show the reaction diagram
-
i.e. angiotensin I, 11.3% degradation after 16 h
-
?
Gly-L-Pro-L-Met-2-anthraquinonyl hydrazide + H2O
Gly-L-Pro-L-Met + 2-anthraquinonyl hydrazine
show the reaction diagram
-
-
-
-
?
Gly-L-Pro-L-Met-4-hydrazino-N-hexyl-1,8-naphthalimide + H2O
?
show the reaction diagram
-
-
-
-
?
Gly-Lys-Pro-Ile-Pro-Phe-Phe-Arg-Leu-Lys + H2O
Gly-Lys-Pro-Ile-Pro-Phe + Phe-Arg-Leu-Lys
show the reaction diagram
-
-
-
-
?
Gly-Pro-Ala 4-nitroanilide + H2O
Gly-Pro-Ala + 4-nitroaniline
show the reaction diagram
-
at 20% the rate of Gly-Pro-Met 4-nitroanilide hydrolysis
-
-
?
Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
at 10% the rate of Gly-Pro-Met 4-methylcoumarin 7-amide hydrolysis
-
-
?
Gly-Pro-Leu 2-naphthylamide + H2O
Gly-Pro-Leu + 2-naphthylamine
show the reaction diagram
-
at 30% the rate of Gly-Pro-Met 2-naphthylamide hydrolysis
-
-
?
Gly-Pro-Leu 2-naphthylamide + H2O
Gly-Pro-Leu + 2-naphthylamine
show the reaction diagram
-
25% of the activity with Gly-Pro-Met 2-naphthylamide, Ala-Ala-Phe-4-nitroanilide or Ala-Ala-Phe-7-amido-4-methylcoumarin
-
-
?
Gly-Pro-Met 2-naphthylamide + H2O
Gly-Pro-Met + 2-naphthylamine
show the reaction diagram
-
-
-
?
Gly-Pro-Met 2-naphthylamide + H2O
Gly-Pro-Met + 2-naphthylamine
show the reaction diagram
-
-
-
-
?
Gly-Pro-Met 2-naphthylamide + H2O
Gly-Pro-Met + 2-naphthylamine
show the reaction diagram
-
-
-
-
?
Gly-Pro-Met 4-nitroanilide + H2O
Gly-Pro-Met + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Gly-Pro-Met-7-amido-4-methylcoumarin + H2O
Gly-Pro-Met + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
GNLWATGHFM-NH2 + H2O
GNL + WATGHFM-NH2
show the reaction diagram
-
i.e. neuromedin B, complete degradation after 16 h
-
?
His-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
His-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Phe-4-hydrazino-N-hexyl-1,8-naphthalimide + H2O
?
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Neuromedin B + H2O
?
show the reaction diagram
-
-
-
-
?
Phe-Pro-Ala 2-naphthylamide + H2O
Phe-Pro-Ala + 2-naphthylamine
show the reaction diagram
-
synthetic peptide modeled after NH2-terminal tripeptide sequence of the phenylalanyl monomer of bovine or rat growth hormone, fluorogenic substrate
-
?
Phe-Pro-Ala-4-nitroanilide + H2O
Phe-Pro-Ala + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedB
-
-
?
Phe-Pro-Ala-4-nitroanilide + H2O
Phe-Pro-Ala + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedC
-
-
?
Phe-Pro-Ala-4-nitroanilide + H2O
Phe-Pro-Ala + 4-nitroaniline
show the reaction diagram
Q70GH4, Q70J58, Q70J59
SedD
-
-
?
phenylalanyl monomer of bovine growth hormone + H2O
?
show the reaction diagram
-
cleaves 11 tripeptides sequentially from the NH2-terminus
-
-
?
poly(Gly-Pro-Ala) + H2O
Gly-Pro-Ala
show the reaction diagram
-
-
-
?
RVYIHPF + H2O
RVY + IHPF
show the reaction diagram
-
i.e. angiotensin III, 95.0% degradation after 15 min
-
?
Ser-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ser-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
show the reaction diagram
-
-
-
-
?
subunit C of mitochondrial ATP synthase + H2O
?
show the reaction diagram
-
-
-
-
?
synthetic collagen-like polypetides + H2O
Gly-Pro-Xaa tripeptides
show the reaction diagram
-
-
-
?
Val-Pro-Arg 4-nitroanilide + H2O
Val-Pro-Arg + 4-nitroaniline
show the reaction diagram
-
at 11% the rate of Gly-Pro-Met 4-nitroanilide hydrolysis
-
-
?
YGGFLRKYP + H2O
YGG + FLRKYP
show the reaction diagram
-
i.e. beta-neo-endorphin, 28% degradation after 16 h
-
?
[Ala-Ala-Phe]2-rhodamine 110 + H2O
[Ala-Ala-Phe]2 + rhodamine 110
show the reaction diagram
-
specific substrate for determining TPP-I activity and intracellular localization in living cells
-
-
?
[Arg-Nle-Nle]2-rhodamine 110 + H2O
[Ala-Ala-Phe]2 + rhodamine 110
show the reaction diagram
-
specific substrate for determining TPP-I activity and intracellular localization in living cells
-
-
?
GKPIPFFRLK + H2O
GKPIP + FFRLK
show the reaction diagram
-
endo-type substrate, 24.5% degradation after 16 h
-
?
glucagon + H2O
additional information
-
-
-
all nine tripeptides and the C-terminal pentapeptide are identified
?
L-Arg-L-Nle-L-Nle-7-amido-4-methylcoumarin + H2O
L-Arg-L-Nle-L-Nle + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
N-terminal exopeptidase that removes tripeptide units provided the P3 residue is unsubstituted
-
-
-
additional information
?
-
-
no significant aminopeptidase activity
-
-
-
additional information
?
-
-
succinyl-Ala-Ala-Phe-7-amido-4-methylcoumarin, succinyl-Gly-Pro-Leu 4-methylcoumarin 7-amide, Gly-Pro-7-amido-4-methylcoumarin, Gly-7-amido-4-methylcoumarin, Pro-7-amido-4-methylcoumarin, Met-7-amido-4-methylcoumarin, Ala 7-amido-4-methylcoumarin, Phe-7-amido-4-methylcoumarin, or Leu-7-amido-4-methylcoumarin
-
-
-
additional information
?
-
-
substrates not cleaved after 24 h of digestion: dynorphin, beta-casomorphin-7, Leu-enkephalin, Met-enkephalin, neurokinin-A, LVV-hemorphin, bradykinin
-
-
-
additional information
?
-
-
the enzyme removes tripeptides from the free N-termini of small polypeptides and also shows a minor endoprotease activity
-
-
-
additional information
?
-
-
Met-2-naphthylamide, succinyl-Gly-Pro-Met 2-naphthylamide, Pro-Met-7-amido-4-methylcoumarin, Met-7-amido-4-methylcoumarin, methoxysuccinyl-Gly-Pro-Met-7-amido-4-methylcoumarin, benzyloxy-Arg-Arg-7-amido-4-methylcoumarin, Arg-7-amido-4-methylcoumarin, Pro-Ala-4-nitroanilide, Ala 4-nitroanilide, tert-butyloxycarbonyl-Gly-Pro-Ala 4-nitroanilide, benzoyl-Val-Pro-Arg 4-nitroanilide
-
-
-
additional information
?
-
-
the enzymne cleaves tripeptides from synthetic substrates provided that the N-terminus is unsubstituted and the amino acid in the P1 position is not charged. The enzyme also cleaves small peptides, angiotensin II and glucagon, releasing tripeptides but does not appear to demonstrate any preference for amino acids on either side of the cleavage site. Substrates with a charged amino acid in the P1 position appear to be resistant to hydrolysis
-
-
-
additional information
?
-
-
absolute requirement for unsubstituted amino-terminus
-
-
-
additional information
?
-
-
no substrates are Ala-Phe-Pro-beta-naphthylamide (modeled after NH2-terminal tripeptide sequence of the alanyl monomer of bovine growth hormone), alanyl monomer of bovine growth hormone
-
-
-
additional information
?
-
-
an inherited deficiency of tripeptidyl peptidase I activity causes a fatal lysosomal storage disorder, classic late infantile neuronal ceroid lipofuscinosis, CLN2
-
-
-
additional information
?
-
-
exopeptidase involved in intracellular (lysosomal) degradation of collagen fibrils
-
-
-
additional information
?
-
-
involved in degradation of bone collagen
-
-
-
additional information
?
-
-
classical late infantile neuronal ceroid lipofuscinosis is an autosomal recessive disease caused by mutations in the CLN2 gene resulting in functional defects of the gene product tripeptidyl-peptidase I. This disease is associated with a progressive neurodegenerative course beginning at the age of two years with developmental stagnation, finally leading to a complete loss of motor function, vision and speech by the age of 10 years
-
-
-
additional information
?
-
-
elevated enzyme activity of tripeptidyl peptidase I and other lysosomal enzymes in Sjoegren's syndrome patients may play a role in tissue damage by accelerated breakdown of glycoproteins in lysosomes
-
-
-
additional information
?
-
-
TPP I is the predominant proteolytic enzyme responsible for the intracellular degradation of neuromedin B. The inability of cells from patients with late-infantile neuronal ceroid lipofuscinosis (CNL2) to degrade neuromedin B and other neuropeptides may contribute to the pathogenesis of the disease
-
-
-
additional information
?
-
-
determination of the substrate specificity of tripeptidyl-peptidase I using combinatorial peptide libraries and development of improved fluorogenic substrates
-
-
-
additional information
?
-
-
prefers Leu, Phe and Nle at the P1 position, whereas Asn, His, Lys, Arg, Ser, Val, Ile, Thr, Gly and Pro are highly unfavored in this position, showing less than 1% of the activity of the best substrates
-
-
-
additional information
?
-
-
TPP I acts preferentially on small, unstructured oligopeptides of less than 5 kDa
-
-
-
additional information
?
-
-
dipeptidyl-peptidase I cannot functionally compensate for the loss of tripeptidyl-peptidase I
-
-
-
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
additional information
?
-
-
an inherited deficiency of tripeptidyl peptidase I activity causes a fatal lysosomal storage disorder, classic late infantile neuronal ceroid lipofuscinosis, CLN2
-
-
-
additional information
?
-
-
exopeptidase involved in intracellular (lysosomal) degradation of collagen fibrils
-
-
-
additional information
?
-
-
involved in degradation of bone collagen
-
-
-
additional information
?
-
-
classical late infantile neuronal ceroid lipofuscinosis is an autosomal recessive disease caused by mutations in the CLN2 gene resulting in functional defects of the gene product tripeptidyl-peptidase I. This disease is associated with a progressive neurodegenerative course beginning at the age of two years with developmental stagnation, finally leading to a complete loss of motor function, vision and speech by the age of 10 years
-
-
-
additional information
?
-
-
elevated enzyme activity of tripeptidyl peptidase I and other lysosomal enzymes in Sjoegren's syndrome patients may play a role in tissue damage by accelerated breakdown of glycoproteins in lysosomes
-
-
-
additional information
?
-
-
TPP I is the predominant proteolytic enzyme responsible for the intracellular degradation of neuromedin B. The inability of cells from patients with late-infantile neuronal ceroid lipofuscinosis (CNL2) to degrade neuromedin B and other neuropeptides may contribute to the pathogenesis of the disease
-
-
-
additional information
?
-
-
dipeptidyl-peptidase I cannot functionally compensate for the loss of tripeptidyl-peptidase I
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
contains one Ca2+ ion
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
-
2-mercaptoethanol
-
0.6 mM, complete inactivation
3,4-Dichloroisocoumarin
-
weak
3,4-Dichloroisocoumarin
-
-
acetylpepstatin
-
1 mM, 14% inhibition
Ala-Ala-Ala-CH2Cl
-
competitive
Ala-Ala-Phe-CH2Cl
-
IC50: 0.06 mM
Ala-Ala-Phe-CH2Cl
-
-
Ala-Ala-Phe-CH2Cl
-
0.001 mM, complete inhibition, competitive
Ala-Ala-Phe-chloromethylketone
-
0.1 mM, complete inhibition
Ala-Ala-Tyr(benzoyl)-CH2Cl
-
competitive
Ala-Ala-Tyr-CH2Cl
-
competitive
Ala-Phe-Ala-CH2Cl
-
competitive
aminoethylbenzylsulfonyl fluoride
-
-
angiotensin II
-
0.3 mM, 22% inhibition
Angiotensin III
-
0.3 mM, 18% inhibition
Aprotinin
-
0.08 mM, 60% loss of activity
bestatin
-
0.01 mM, 11% inhibition
chymostatin
-
1 mM, 47% inhibition
Dextran sulfate
-
initially, as the glycosaminoglycan concentration progressed from 0 to about 100 ng/ml, the activity of TPP I falls down rapidly to reach a plateau with saturable inhibition at about 100 ng-0.1 mg/ml and then it gradually increases with increased glycosaminoglycan concentration. The maximal percent inhibition is 50%
-
diethyl dicarbonate
-
61% inhibition with 1 mM, 91% inhibition with 10 mM
diethyldicarbonate
-
-
diisopropylfluorophosphate
-
at high concentration
diisopropylfluorophosphate
-
IC50: 1.15 mM
diisopropylfluorophosphate
-
-
Diprotin A
-
0.1 mM, 50% inhibition
dithiothreitol
-
0.6 mM, complete inactivation
E-64
-
0.01 mM, 22% inhibition
glutathione
-
0.6 mM, 93% loss of activity
Gly-Ala-Ala
-
0.3 mM, 36% inhibition
Gly-His-Lys
-
0.3 mM, 49% inhibition
heparin
-
initially, as the glycosaminoglycan concentration progressed from 0 to about 100 ng/ml, the activity of TPP I falls down rapidly to reach a plateau with saturable inhibition at about 100 ng-0.1 mg/ml and then it gradually increases with increased glycosaminoglycan concentration. The maximal percent inhibition is 50%
Hg2+
-
1 mM, complete inhibition, IC50: 0.67 mM
L-Ala-L-Ala-L-Phe-chloromethyl ketone
-
TPP I specific inhibitor
Leu-Ala-Pro
-
0.3 mM, 55% inhibition
Leupeptin
-
0.001 mg/ml, 17% inhibition
N-acetyl-Ile-Ala-phenylalaninal
-
1 mM, 88% inhibition
N-acetyl-Ile-Pro-phenylalaninal
-
1 mM, 38% inhibition
NaCl
-
2 M, pH 5.0, 22% decrease in TPP I activity
p-chloromercuribenzene sulfonic acid
-
IC50: 0.27 mM
p-tosyl-L-Lys chloromethyl ketone
-
weak
Pepstatin
-
0.1 mM, 15% inhibition
pepstatin A
-
0.1 mg/ml, 26% inhibition
Phe-Ala-Ala-CH2Cl
-
competitive
Phe-CH2Cl
-
0.1 mM, 21% inhibition
-
Phenyl-N-phenylphosphoamidochloridate
-
weak
phenylmethylsulfonyl fluoride
-
weak
prosegment of tripeptidyl peptidase I
-
wild type prosegment of tripeptidyl peptidase I (176 amino acid residues) is a potent, slow-binding inhibitor of its parent enzyme with an overall inhibition constant in the low nanomolar range, in the presence of the prosegment (at 0.001 mM) leads from a significant to complete inhibition of TPP I activity at pH between 3.5 and 5.5 with IC 50 ranging from more than 0.016 mM to 0.000043 mM at pH from 3.5 to 5.5
-
Tyrostatin
-
1 mM, 19% inhibition
Lys-Ala-Ala
-
0.3 mM, 34% inhibition
additional information
-
no inhibition by pepstatin
-
additional information
-
no inhibition: alkali-inactivated diisopropylfluoride, leupeptin; no inhibition by pepstatin; no inhibition: EDTA
-
additional information
-
no inhibition by pepstatin; no inhibition: EDTA; no inhibition: Phe-Pro-Arg chloromethylketone, inhibitors of cysteine proteinases or metalloproteinases, iodoacetamide, trans-epoxysuccinyl-L-leucinamido(4-guanidino)butane (i.e. E-64), mercaptoethanol, bestatin, soybean trypsin inhibitor, phenylchloromethyl ketone
-
additional information
-
resistant to inhibition by diisopropylfluorophosphate
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
neonatally treated mice display high levels of TPP-I activity in the CNS 1 year after administration of AAVrh.10hCLN2 vector directly to the neonatal brain of CLN2 knockout mice at 2 days, 3 weeks, and 7 weeks of age, the AAVrh.10hCLN2 vector contains an expression cassette consisting of the human CLN2 cDNA driven by a CMV/beta-actin hybrid promoter consisting of the enhancer from the cytomegalovirus immediate early gene, the promoter, splice donor and intron from the chicken beta-actin gene, the splice acceptor from the rabbit beta-globin gene followed by a CLN2 cDNA with an optimized Kozak translation initiation sequence, the cDNA is followed by the polyadenylation sequence from rabbit beta-globin, the expression cassette is surrounded by the inverted terminal repeats of adeno-associated virus 2
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.233
-
Ala-Ala-Phe-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.076
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme S475L
0.098
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, wild-type enzyme
0.105
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D276A
0.111
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D360A
0.118
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme E272A
0.164
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D327A
0.188
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 4.5, 30C
0.68
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 4.0
0.73
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 4.0
0.0268
-
Ala-Ala-Phe-p-nitroanilide
-
pH 4.5
0.0148
-
Ala-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.888
-
Ala-Ala-Pro-p-nitroanilide
-
pH 4.5
0.00402
-
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.123
-
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, mutant enzyme E77A
0.00509
-
Ala-Asp-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.0422
-
Ala-His-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.028
-
Ala-Nle-Leu-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.037
-
Ala-Nle-Nle-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.03
-
Ala-Nva-Nle-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.437
-
Ala-Phe-Pro-p-nitroanilide
-
pH 4.5
0.0483
-
Ala-Ser-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.0193
-
Arg-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.054
-
Arg-Pro-Phe-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.11
-
Asp-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.45
-
Gly-Pro-Met 2-naphthylamide
-
-
0.167
-
His-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.0625
-
Phe-Pro-Ala-4-nitroanilide
Q70GH4, Q70J58, Q70J59
pH 6.0, 20C, SedB
0.108
-
Ser-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
5.3
-
Ala-Ala-Phe-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
0.04
1.97
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D276A
0.12
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme S475L
1.52
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D360A
1.54
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme E272A
3.22
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, wild-type enzyme
4.22
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D327A
5.2
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 4.5, 30C
9.36
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, mutant enzyme D276A
33.1
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 4.0
41.66
-
Ala-Ala-Phe-7-amido-4-methylcoumarin
-
pH 5.0, 37C, wild-type enzyme
550000
-
Ala-Ala-Phe-p-nitroanilide
-
pH 4.5
34.5
-
Ala-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
210000
-
Ala-Ala-Pro-p-nitroanilide
-
pH 4.5
0.0124
-
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 378C, mutant enzyme E77A
11.8
-
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
11
-
Ala-Asp-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
26.4
-
Ala-His-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
3.6
-
Ala-Nle-Leu-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
4.2
-
Ala-Nle-Nle-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
4.7
-
Ala-Nva-Nle-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
140000
-
Ala-Phe-Pro-p-nitroanilide
-
pH 4.5
30.7
-
Ala-Ser-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
0.91
-
Arg-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
8.2
-
Arg-Pro-Phe-7-amido-4-carbamoylmethylcoumarin
-
pH 4.5, 30C
18.5
-
Asp-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
13.7
-
His-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
35
-
Phe-Pro-Ala-4-nitroanilide
Q70GH4, Q70J58, Q70J59
pH 6.0, 20C, SedB
4.7
-
Ser-Ala-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37C, wild-type enzyme
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0069
-
Ala-Ala-Ala-CH2Cl
-
pH 4.0, 37C
0.000063
-
Ala-Ala-Phe-CH2Cl
-
pH 4.0, 37C
0.00081
-
Ala-Ala-Tyr(benzoyl)-CH2Cl
-
pH 4.0, 37C
0.00072
-
Ala-Ala-Tyr-CH2Cl
-
pH 4.0, 37C
0.0079
-
Ala-Phe-Ala-CH2Cl
-
pH 4.0, 37C
0.0013
-
Phe-Ala-Ala-CH2Cl
-
pH 4.0, 37C
0.0001581
-
prosegment of tripeptidyl peptidase I
-
in sodium acetate buffer (50 mM, pH 5.0), 0.05% Triton X-100
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.06
-
Ala-Ala-Phe-CH2Cl
-
IC50: 0.06 mM
1.15
-
diisopropylfluorophosphate
-
IC50: 1.15 mM
0.67
-
Hg2+
-
1 mM, complete inhibition, IC50: 0.67 mM
0.27
-
p-chloromercuribenzene sulfonic acid
-
IC50: 0.27 mM
0.0001292
-
prosegment of tripeptidyl peptidase I
-
in sodium acetate buffer (50 mM, pH 5.0), 0.05% Triton X-100
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.516
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
-
-
assay at
4
-
-
-
4
-
-
hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin
4.3
-
-
hydrolysis of Gly-Pro-Met 4-nitroanilide
4.5
5
-
hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin
4.5
-
-
hydrolysis of Gly-Pro-Met 2-naphthylamide
4.5
-
-
and a second optimum at pH 5.9
5
5.5
-
hydrolysis of Gly-Pro-Met 2-naphthylamide or Ala-Ala-Phe 4-methylcoumarin 7-amide
5
-
-
hydrolysis of Gly-Pro-Met 4-methylcoumarin 7-amide
5
-
Q70GH4, Q70J58, Q70J59
SedD
5.9
-
-
and a second optimum at pH 4.5, at pH 5.9 70% of the activity at pH 4.5
6
-
Q70GH4, Q70J58, Q70J59
SedB; SedC
additional information
-
-
the optimum pH of TPP 1 is dependent on the substrate used
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1.5
6
Q70GH4, Q70J58, Q70J59
active between pH 3 and pH 7, SedD
3
4.8
-
pH 3.0: about 30% of maximal activity, pH 4.8: about 50% of maximal activity
3
7
Q70GH4, Q70J58, Q70J59
active between pH 3 and pH 7, SedB; active between pH 3 and pH 7, SedC
3
8
-
about half-maximal activity at pH 3 and 8, hydrolysis of Gly-Pro-Met 2-naphthylamide
3.5
5
-
pH 3.5: about 25% of maximal activity, pH 5.0: about 45% of maximal activity, hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin
3.5
6.5
-
about half-maximal activity at pH 3.5 and 6.5, hydrolysis of Ala-Ala-Phe 4-methylcoumarin 7-amide
4
5.5
-
about 55% of maximal activity at pH 4.0 and at pH 5.5, hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin
37
-
-
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.12
-
Q70GH4, Q70J58, Q70J59
calculated from sequence, mature domain, SedD
5.17
-
Q70GH4, Q70J58, Q70J59
calculated from sequence, mature domain, SedB
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
TPPI is expressed ubiquitously throughout the body but disease appears restricted to the brain
Manually annotated by BRENDA team
-
activity in neuroglia cells, no activity in small granule cells, Purkinje cells, remified dendrites and myelinated fibers
Manually annotated by BRENDA team
-
activity in proximal tubules, distal tubules, collecting tubules and transitional cells, no activity in glomerulus and Bowmans capsule
Manually annotated by BRENDA team
-
highest activity in kidney
Manually annotated by BRENDA team
-
enzyme activity is elevated during the first 5 years of Sjoegren's syndrome, and it increases further between 5 and 10 years after diagnosis
Manually annotated by BRENDA team
-
activity in corpus luteum and corpus albicus, no activity in follicles
Manually annotated by BRENDA team
-
activity in ducts, no activity in acinar cells and islets of Langerhans
Manually annotated by BRENDA team
-
activity in excretory duct and stereocilia, no activity in glandular acinus and intercalated portion
Manually annotated by BRENDA team
-
activity in red pulp, no activity in white pulp
Manually annotated by BRENDA team
-
heterogenous distribution of TPP I positive macrophages, leukocytes and reticular fibroblasts in the red and white pulp
Manually annotated by BRENDA team
-
activity in spermatogonia, spermatozoa, Sertoli cells and Leygig cells, no activity in basement cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
the conversion of the proenzyme into the mature form takes place in lysosomal compartment
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22000
-
-
TPP I prosegment, SDS-PAGE
46000
-
-
mature protein, SDS-PAGE
46000
-
-
SDS-PAGE
46000
-
-
autoprocessed TPP1, SDS-PAGE
46000
-
-
SDS-PAGE, processed form
48000
-
-
mature enzyme, SDS-PAGE
55000
-
-
gel filtration in the presence of 3 M urea
57000
-
-
-
64000
-
-
gel filtration
66000
-
-
immature protein, SDS-PAGE
68000
-
-
pro-enzyme, SDS-PAGE
68000
-
-
SDS-PAGE, proenzyme
217000
-
-
gel filtration
250000
-
-
enzyme aggregate, gel filtration
280000
-
-
non-denaturing PAGE
290000
-
-
gel filtration
700000
-
-
gel filtration in the absence or presence of 4 M urea
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 48000, SDS-PAGE
?
-
x * 46000, SDS-PAGE in presence of 2-mercaptoethanol
?
Q70GH4, Q70J58, Q70J59
x * 40600, SedD, calculated from sequence; x * 43000, SedB, calculated from sequence
?
-
x * 46000, SDS-PAGE
dimer
-
2 * 107000, SDS-PAGE
monomer
-
1 * 47000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
Q70GH4, Q70J58, Q70J59
the preproprotein SedB is composed of 602 amino acids, the signal peptide contains 24 amino acids; the preproprotein SedC is composed of 596 amino acids, the signal peptide contains 24 amino acids; the preproprotein SedD is composed of 568 amino acids, the signal peptide contains 20 amino acids
glycoprotein
-
all five potential N-glycosylation sites in the enzyme are utilized, some of its N-linked oligosaccharides are of the complex/hybrid type
glycoprotein
-
TPP I in vivo utilizes all five N-glycosylation sites (Asn210, Asn222, Asn286, Asn313 and Asn443). Mutation of N286Q dramatically affects the folding of the enzyme and partially arrests TPP I in the endoplasmic reticulum. N-glycans at Asn210, Asn222, Asn313, and Asn443 contribute slightly to the specific activity of the enzyme
glycoprotein
-
TPP 1 is glycosylatd at most or all of five asparagin-linked glycosylation sites
glycoprotein
-
proteolytic modification
glycoprotein
-
TPP1 requires at least partial glycosylation for in vitro autoprocessing and proteolytic activity
glycoprotein
-
-
proteolytic modification
-
the 68000 Da precursor polypeptide is converted to the mature enzyme of 48000 Da. Although tripeptidyl-peptidase I zymogen is capable of autoactivation in vitro, a serine protease that is sensitive to 4-(2-aminoethyl)-benzene-sulfonamide participates in processing of the proenzyme to the mature, active form in vivo
proteolytic modification
-
the protein is synthesized as an inactive zymogen that is autocatalytically converted to an active serine protease at acidic pH, removal of a 19-residue signal peptide
proteolytic modification
-
TPP-I is synthesized as a 66000 Da inactive precursor consisting of a 195 residue prepro-peptide and 368 residue mature enzyme. Most of the precursor proteins are converted to a 46000 Da form by post-translational proteolysis
proteolytic modification
-
proenzyme, the N-linked glycan at Asn286 is involved in maturation of the enzyme
proteolytic modification
-
TPP I activity is not involved in the major proteolytic step leading to generation of the mature enzyme, indicating that autoactivation of human pro-TTP I in vitro occurs by intramolecular mechanisms
proteolytic modification
-
autoactivation of pro-TPP I is fully activated at pH 3.5 after only 15 min of activation in 50 mM sodium acetate buffer, independent of NaCl concentration. In presence of 1 M NaCl (but not 150 mM NaCl) significant activation also is observed at pH 4.0. Ionic strength affects the yield but not the rates or the mechanism of pro-TPP I activation in vitro
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
deglycosylated inactive proenzyme pro-TPP1, hanging drop vapour diffusion method, using 0.1 M citrate, 4-7% polyethylene glycol 6000, at pH 5.0
-
fully-glycosylated TPP1 precursor, hanging drop vapour diffusion method, using 7% PEG4000, 0.02 M zinc sulfate, 0.1 M sodium acetate, pH 5.0, and 0.1 M ammonium sulfate
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2
5
-
optimal activation occurs in the range pH 3.0-4.0, the proteolytic processing takes place in a wider pH range (2.0-5.0), at pH 3.0 and lower, TPP I is quickly inactivated, whereas the polypeptide generated at higher pH (4.5-5.0) possesses 6- and 13-aa N-terminal extensions and is inactive, the enzyme is unstable at alkaline and neutral pH
2.5
5
-
stable
3
6
-
at pH 3.0 the activity of TPP I is less than 75% of its maximal activity at pH 4.5, most stable at pH 3-4, no activity at pH above 6.0
3
-
-
has weak endoproteolytic activity at pH 3
3.5
4.5
-
stable
3.5
-
-
TPP I is an acidic protease that is quickly inactivated under alkaline pH conditions, in the absence of prosegment, the enzyme is quickly denatured with a rate constant of around 0.107/min, amounting to a half-life of 6.472 min, whereas in the presence of the prosegment, the inactivation rate is reduced to about 0.035/min, and approximately 84% of the activity is preserved after over 2 h of incubation
5
6
-
48 h, stable
7
-
-
48 h, unstable above
7.4
-
-
37C, half-life: of mature enzyme in absence of heparin is 2.5 min, of mature enzyme in presence of heparin is 21.5 min
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
binding to glycosaminoglycans improves the thermal stability of TPP I
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glycosyminoglycans protect against alkaline pH-induced denaturation
-
resistant against freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, purified pro-TPP I is stable in 50 mM Tris buffer in 50% glycerol
-
4C, pH 4.0, 50 mM sodium acetate buffer, stable for 10-14 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant SedB; recombinant SedD
Q70GH4, Q70J58, Q70J59
butyl-Sepharose 4 column chromatography, Mono Q12 column chromatography, and Superdex 75 gel filtration
-
HisTrap HP column chromatography
-
Ni2+ affinity resin chromatography and heparin agarose affinity chromatography
-
recombinant enzyme
-
Sepharose 4 FF column chromatography and Blue Sepharose column chromatography
-
partial
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of SedB in Pichia pastoris; expression of SedC in Pichia pastoris; expression of SedD in Pichia pastoris
Q70GH4, Q70J58, Q70J59
C-terminal hexahistidine-tagged human CLN2p/tripeptidyl-peptidase I produced from insect cells transfected with a baculovirus vector
-
expressed in Chinese hamster ovary cells
-
expressed in CHO cells
-
expressed in HEK-293 cells
-
expressed in Mus musculus brain
O14773
expressed in Rattus norvegicus brain
-
expression in Chinese hamster ovary cells under the control of the cytomegalovirus promoter
-
expression in silkworm pupae
-
expression of mutant enzymes in HEK cells and CHO cells
-
expression of mutant enzymes N210Q, N222Q, N286Q, N313Q and N443N in Chinese hamster overy cells and human embryonic kidney 293 cells
-
expression of wild-type and mutant enzymes (E272A, D276A, D327A, D360A and S475L) in CHO cells
-
TPP I prosegment is expressed in Escherichia coli BL21 (DE3) cells
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C365R
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decreased activity
C365R
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
D165A
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inactive mutant
D276A
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kcat/Km is 21% of the wild-type value
D327A
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kcat/Km is 6% of the wild-type value
D360A
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lack of enzyme activity and processing
D360A
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kcat/Km is 3% of the wild-type value
D517A
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lack of enzyme activity and processing
D81A
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not expressed in Sf9 cells
E272A
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kcat/Km is 3% of the wild-type value
E343K
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
E343L
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decreased activity
E77A
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very low activity with Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
G284V
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decreased activity
G284V
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the mutation probably compromises the active center and results in loss of proteolytic activity
G284V
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
G473R
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the mutation probably compromises the active center and results in loss of proteolytic activity
G77R
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decreased activity
G77R
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the mutation is associated with classic late infantile neuronal ceroid lipofuscinosis
G77R
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, 1% of wild-type activity
I287N
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mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 4.1% of wild-type enzyme when expressed in HEK cells, blocked processing to mature size peptidase leads to protein retention in the endoplasmic reticulum and rapid degradation in non-lysosomal compartments
I287N
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decreased activity
I287N
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
K428N
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no apparent conformational destabilization is observed for the missense mutation
N286Q
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the secreted proenzyme formes non-native, interchain disulfide bridges and displays only residual TPP I activity upon acidification. A small portion of the mutant enzyme reaches the lysosome and is processed to an active species, however, it shows low thermal and pH stability
N286S
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mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 5.8% of wild-type enzyme when expressed in HEK cells, blocked processing to mature size peptidase leads to protein retention in the endoplasmic reticulum and rapid degradation in non-lysosomal compartments
N286S
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decreased activity
N286S
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the substitution results in loss of one glycosylation site, which leads to almost complete loss of protease activity
N286S
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
P202L
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decreased activity
P202L
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
P544S
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decreased activity; demonstrates a normal polypeptide pattern on Western blots, enzyme activity, and lysosomal localization
P544S
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protein processing similar to wild-type, lysosomal localisation, 32.8% of wild-type activity
Q248P
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the mutation probably compromises the active center and results in loss of proteolytic activity
Q422H
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mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 4.7% of wild-type enzyme when expressed in HEK cells, blocked processing to mature size peptidase leads to protein retention in the endoplasmic reticulum and rapid degradation in non-lysosomal compartments
Q422H
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decreased activity
Q442H
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
R127Q
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mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 74.3% of wild-type enzyme when expressed in HEK cells
R127Q
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decreased activity; demonstrates a normal polypeptide pattern on Western blots, enzyme activity, and lysosomal localization
R127Q
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protein processing similar to wild-type,lysosomal localization, 43% of wild-type activity
R206C
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decreased activity
R206C
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, 0.7% of wild-type activity
R208X
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mutation is identified in patients with late infantile ceroid lipofuscinosis, no detection of any translational product for the mutant
R266Q
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no apparent conformational destabilization is observed for the missense mutation
R447H
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decreased activity
R447H
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, 1.8% of wild-type activity
S280A
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inactive mutant
S475
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inactive mutant enzyme
S475L
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kcat/Km is 0.4% of the wild-type value
S475L
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decreased activity; inactive
S475L
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the mutation probably compromises the active center and results in loss of proteolytic activity
S475L
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protein processing similar to wild-type, lysosomal localization, no enzymatic activity
T353P
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mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 5.5% of wild-type enzyme when expressed in HEK cells, blocked processing to mature size peptidase leads to protein retention in the endoplasmic reticulum and rapid degradation in non-lysosomal compartments
V216M
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no apparent conformational destabilization is observed for the missense mutation
V227M
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protein processing different from wild-type, mutant is not localized in lysosomes, intracellular trafficking of mutant is altered compared to wild-type, no enzymatic activity
V277M
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decreased activity
V277M
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the mutation probably compromises the active center and results in loss of proteolytic activity
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
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adeno-associated virus 2-mediated CLN2 gene transfer to rodent and non-human primate brain results in long-term TPP-I expression compatible with therapy for late infantile neuronal ceroid lipofuscinosis
diagnostics
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potential markers for Sjoegren's syndrome
diagnostics
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saliva is a reliable and non-invasive source for the diagnosis of infantile (CLN1) and late infantile (CLN2) neuronal ceroid lipofuscinoses
medicine
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[Ala-Ala-Phe]2-rhodamine 110 and [Arg-Nle-Nle]2-rhodamine 110 are specific substrate for determining TPP-I activity and intracellular localization in living cells. These substrates can be a valuable tool for studying the neuronal pathology underlying classical late-infantile neuronal ceroid lipofuscinosis
medicine
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mutations in tripeptidyl-peptidase I underlie the classic late-infantile form of neuronal ceroid lipofuscinoses (CLN2), the most common neurodegenerative disorders of childhood
molecular biology
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fluorescent method for the histochemical detection of tripeptidyl peptidase I using glycyl-L-prolyl-L-Met-2-anthraquinonyl hydrazide as substrate