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Information on EC 3.4.14.9 - tripeptidyl-peptidase I and Organism(s) Homo sapiens and UniProt Accession O14773
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3.4.14.9 tripeptidyl-peptidase ISpecify your search results
Word Map
- 3.4.14.9
-
infantile
- neurodegenerative
-
lincl
-
late-infantile
- lipofuscinoses
- batten
-
curvilinear
- palmitoyl-protein
-
pepstatin-insensitive
- cdc28p
-
serine-carboxyl
-
lipopigments
- molecular biology
- diagnostics
- medicine
- analysis
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
Release of an N-terminal tripeptide from a polypeptide, but also has endopeptidase activity
Synonyms
tpp-i, tripeptidyl peptidase 1, cln2p, tripeptidyl peptidase i, tripeptidyl-peptidase i, cln2 protein, tripeptidyl-peptidase 1, tripeptidyl peptidase-i, tpp1f, ttp-i, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Tpp1
aminopeptidase, tripeptidyl, I
-
-
-
-
LPIC
-
-
-
-
lysosomal pepstatin insensitive protease
-
-
-
-
TPP I
tripeptidyl aminopeptidase
-
-
-
-
tripeptidyl aminopeptidase I
-
-
-
-
tripeptidyl peptidase
-
-
-
-
tripeptidyl peptidase I
Tpp1
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
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
tripeptidyl peptidase I
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
151662-36-1
-
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
amyloid-beta + H2O
?
AbetaCy3 peptides are released from the nanofibrils due to TPP1 activity
-
-
?
amyloid-beta1-42 + H2O
?
the 34 Abeta end of the substrate shows integrated peak areas for peptide fragments 21-34, 22-34, and 23-34, indicative of cleavage after residue L34. The most abundant cleavages occur after residues Y10, G33, L34, and A30, and these cleavages occur more rapidly at pH 3.0 than at pH 4.5, consistent with endopeptidase activity. Peptides ending at residues E11, L17, F20, G37, and G38, are detected with lower abundances. At later times, the abundance of some of the peptides may decrease due to further proteolysis by TPP1. TPP1 can proteolyze monomeric Abeta1-42 efficiently at acidic pH
-
-
?
epsilon-aminocaproyl-WFFIQ-[N-(2,4-dinitrophenyl)-ethylenediamine] + H2O
epsilon-aminocaproyl-WF + FIQ-[N-(2,4-dinitrophenyl)-ethylenediamine]
-
-
-
?
KWFFIQ-[N-(2,4-dinitrophenyl)-ethylenediamine] + H2O
KWF + FIQ-[N-(2,4-dinitrophenyl)-ethylenediamine]
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
L-Arg-L-Nle-L-Nle-7-amido-4-methylcoumarin + H2O
L-Arg-L-Nle-L-Nle + 7-amino-4-methylcoumarin
-
-
-
?
RWFFIQ-[N-(2,4-dinitrophenyl)-ethylenediamine] + H2O
RWF + FIQ-[N-(2,4-dinitrophenyl)-ethylenediamine]
FRET substrate
-
-
?
RWFVIQ-[N-(2,4-dinitrophenyl)-ethylenediamine] + H2O
RWF + VIQ-[N-(2,4-dinitrophenyl)-ethylenediamine]
-
-
-
?
Ala-Ala-Phe-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-carbamoylmethylcoumarin
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
Ala-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Arg-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Ala-Asp-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Asp-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Ala-His-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-His-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Ala-Nle-Leu-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nle-Leu + 7-amino-4-carbamoylmethylcoumarin
-
-
-
-
?
Ala-Nle-Nle-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nle-Nle + 7-amino-4-carbamoylmethylcoumarin
-
-
-
-
?
Ala-Nva-Nle-7-amido-4-carbamoylmethylcoumarin + H2O
Ala-Nva-Nle + 7-amino-4-carbamoylmethylcoumarin
-
-
-
-
?
Ala-Ser-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ala-Ser-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Arg-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Arg-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
Arg-Pro-Phe-7-amido-4-carbamoylmethylcoumarin + H2O
Arg-Pro-Phe + 7-amino-4-carbamoylmethylcoumarin
-
-
-
-
?
Asp-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Asp-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
His-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
His-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
Ser-Ala-Phe-p-nitrophenylalanyl-Arg-Leu + H2O
Ser-Ala-Phe + p-nitrophenylalanyl-Arg-Leu
-
-
-
-
?
synthetic collagen-like polypetides + H2O
Gly-Pro-Xaa tripeptides
-
-
-
?
[Ala-Ala-Phe]2-rhodamine 110 + H2O
[Ala-Ala-Phe]2 + rhodamine 110
-
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
-
specific substrate for determining TPP-I activity and intracellular localization in living cells
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
-
-
-
?
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
L-Ala-L-Ala-L-Phe + 7-amino-4-methylcoumarin
-
-
-
-
?
additional information
?
-
validation and evaluation of a rapid and simple fluorometric tripeptidyl peptidase 1 (TPP1) assay using dried blood specimens to diagnose CLN2 disease
-
-
?
additional information
?
-
-
validation and evaluation of a rapid and simple fluorometric tripeptidyl peptidase 1 (TPP1) assay using dried blood specimens to diagnose CLN2 disease
-
-
?
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
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-
?
additional information
?
-
-
the enzyme removes tripeptides from the free N-termini of small polypeptides and also shows a minor endoprotease activity
-
-
?
additional information
?
-
-
absolute requirement for unsubstituted amino-terminus
-
-
?
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
-
-
?
NATURAL SUBSTRATE
NATURAL 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
amyloid-beta + H2O
?
AbetaCy3 peptides are released from the nanofibrils due to TPP1 activity
-
-
?
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
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ala-Ala-Phe-chloromethylketone
AAF-CMK, inhibits both the endoproteolytic and tripeptidyl peptidase activities of TPP1
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%
-
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%
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
-
additional information
-
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
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00509
Ala-Asp-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37°C, wild-type enzyme
0.076
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme S475L
0.098
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, wild-type enzyme
0.105
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D276A
0.111
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D360A
0.118
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme E272A
0.164
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D327A
0.00402
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37°C, wild-type enzyme
0.123
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 37°C, mutant enzyme E77A
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.04 - 1.97
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D276A
0.12
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme S475L
1.52
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D360A
1.54
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme E272A
4.22
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D327A
9.36
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, mutant enzyme D276A
41.66
Ala-Ala-Phe-7-amido-4-methylcoumarin
pH 5.0, 37°C, wild-type enzyme
0.0124
Ala-Arg-Phe-p-nitrophenylalanyl-Arg-Leu
-
pH 4.0, 378°C, mutant enzyme E77A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0001581
prosegment of tripeptidyl peptidase I
-
in sodium acetate buffer (50 mM, pH 5.0), 0.05% Triton X-100
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0001292
prosegment of tripeptidyl peptidase I
Homo sapiens
-
in sodium acetate buffer (50 mM, pH 5.0), 0.05% Triton X-100
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 5.5
-
hydrolysis of Gly-Pro-Met 2-naphthylamide or Ala-Ala-Phe 4-methylcoumarin 7-amide
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
3 - 4.8
-
pH 3.0: about 30% of maximal activity, pH 4.8: about 50% of maximal activity
3 - 8
-
about half-maximal activity at pH 3 and 8, hydrolysis of Gly-Pro-Met 2-naphthylamide
3.5 - 6.5
-
about half-maximal activity at pH 3.5 and 6.5, hydrolysis of Ala-Ala-Phe 4-methylcoumarin 7-amide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
enzyme activity is elevated during the first 5 years of Sjoegren's syndrome, and it increases further between 5 and 10 years after diagnosis
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
-
-
the conversion of the proenzyme into the mature form takes place in lysosomal compartment
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
CLN2 disease is a genetic disorder caused by dysfunction of the lysosomal enzyme tripeptidyl peptidase 1 (TPP1) that belongs to the neuronal ceroid lipofuscinoses (NCL) and leads to epilepsy, dementia, and death in young persons. CLN2 disease has become treatable by enzyme replacement, which can only be effective when the disease is diagnosed early. Analysis of reliability of a test for TPP1 deficiency in dried blood specimens (DBS) to detect CLN2 disease, overview. Diminished TTP1 activity is carefully checked for clinical information compatible with the diagnosis of CLN2 disease, and respective patients are subject to molecular genetic testing and confirmation of CLN2 disease by detection of known variants within the CLN2 gene, phenotypes, overview
physiological function
tripeptidyl peptidase 1 (TPP1) is a lysosomal serine protease, that possesses endopeptidase activity and cleaves peptides between hydrophobic residues. TPP1 is able to proteolyze fibrillar amyloid-beta efficiently. Mass spectrometry analysis of peptides released from fibrillar amyloid-beta digested with TPP1 reveals several endoproteolytic cleavages including some within beta-sheet regions that are important for fibril formation. These cleavages destabilize fibrillar beta-sheet structure. N-terminal tripeptidyl exopeptidase activity with a pH optimum of 5 that catalyzes the sequential release of tripeptides from the unsubstituted N termini of proteins
additional information
molecular dynamics (MD) simulations are used to analyze the effects of each cleavage on beta-sheet and fibril stability of amyloid-beta, eight cleavage sites are selected to be simulated, namely after residues K16, F20, G33, L34, M35, V36, G38, and V40, stability of hydrogen bonds following selected TPP1 cleavages and peptide release from the fibril, molecular modeling, detailed overview
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). TPP1_HUMAN
563
0
61248
Swiss-Prot
Secretory Pathway (Reliability: 1)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
46000
46000
48000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
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
glycoprotein
proteolytic modification
glycoprotein
TPP1 requires at least partial glycosylation for in vitro autoprocessing and proteolytic activity
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
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
-
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
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
-
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
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C365R
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
D360A
E343K
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
G284V
G473R
the mutation probably compromises the active center and results in loss of proteolytic activity
G77R
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
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
no apparent conformational destabilization is observed for the missense mutation
N286S
P202L
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
protein processing similar to wild-type, lysosomal localisation, 32.8% of wild-type activity
Q248P
the mutation probably compromises the active center and results in loss of proteolytic activity
Q442H
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
protein processing similar to wild-type,lysosomal localization, 43% of wild-type activity
R206C
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
R266Q
no apparent conformational destabilization is observed for the missense mutation
R447H
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
S475L
V216M
no apparent conformational destabilization is observed for the missense mutation
V227M
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
the mutation probably compromises the active center and results in loss of proteolytic activity
G77R
I287N
N286Q
-
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
P544S
Q422H
R127Q
R208X
-
mutation is identified in patients with late infantile ceroid lipofuscinosis, no detection of any translational product for the mutant
S475L
T353P
-
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
G284V
the mutation probably compromises the active center and results in loss of proteolytic activity
G284V
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
N286S
the substitution results in loss of one glycosylation site, which leads to almost complete loss of protease activity
N286S
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
S475L
the mutation probably compromises the active center and results in loss of proteolytic activity
S475L
protein processing similar to wild-type, lysosomal localization, no enzymatic activity
G77R
-
the mutation is associated with classic late infantile neuronal ceroid lipofuscinosis
I287N
-
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
N286S
-
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
P544S
-
demonstrates a normal polypeptide pattern on Western blots, enzyme activity, and lysosomal localization
Q422H
-
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
R127Q
-
mutation is identified in patients with late infantile ceroid lipofuscinosis, enzyme shows 74.3% of wild-type enzyme when expressed in HEK cells
R127Q
-
demonstrates a normal polypeptide pattern on Western blots, enzyme activity, and lysosomal localization
pH STABILITY
ORGANISM
UNIPROT
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
678513
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
698756
7.4
-
37°C, half-life: of mature enzyme in absence of heparin is 2.5 min, of mature enzyme in presence of heparin is 21.5 min
665697
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
binding to glycosaminoglycans improves the thermal stability of TPP I
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
butyl-Sepharose 4 column chromatography, Mono Q12 column chromatography, and Superdex 75 gel filtration
Ni2+ affinity resin chromatography and heparin agarose affinity chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
C-terminal hexahistidine-tagged human CLN2p/tripeptidyl-peptidase I produced from insect cells transfected with a baculovirus vector
expressed in CHO cells
expression of wild-type and mutant enzymes (E272A, D276A, D327A, D360A and S475L) in CHO cells
expression in Chinese hamster ovary cells under the control of the cytomegalovirus promoter
-
expression of mutant enzymes N210Q, N222Q, N286Q, N313Q and N443N in Chinese hamster overy cells and human embryonic kidney 293 cells
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
development of fluorescence resonance energy transfer peptides using tryptophan as the fluorophore to study TPP-I hydrolytic properties. Assay can be applied to spleen and kidney homogenate
diagnostics
medicine
diagnostics
-
saliva is a reliable and non-invasive source for the diagnosis of infantile (CLN1) and late infantile (CLN2) neuronal ceroid lipofuscinoses
medicine
-
[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
-
mutations in tripeptidyl-peptidase I underlie the classic late-infantile form of neuronal ceroid lipofuscinoses (CLN2), the most common neurodegenerative disorders of childhood
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Page, A.E.; Fuller, K.; Chambers, T.J.; Warburton, M.J.
Purification and characterization of a tripeptidyl peptidase I from human osteoclastomas: evidence for its role in bone resorption
Arch. Biochem. Biophys.
306
354-359
1993
Homo sapiens
Golabek, A.A.; Kida, E.; Walus, M.; Wujek, P.; Mehta, P.; Wisniewski, K.E.
Biosynthesis, glycosylation, and enzymatic processing in vivo of human tripeptidyl-peptidase I
J. Biol. Chem.
278
7135-7145
2003
Homo sapiens
Lin, L.; Sohar, I.; Lackland, H.; Lobel, P.
The human CLN2 protein/tripeptidyl-peptidase I is a serine protease that autoactivates at acidic pH
J. Biol. Chem.
276
2249-2255
2001
Homo sapiens (O14773), Homo sapiens
Kopan, S.; Sivasubramaniam, U.; Warburton, M.J.
The lysosomal degradation of neuromedin B is dependent on tripeptidyl peptidase-I: evidence for the impairment of neuropeptide degradation in late-infantile neuronal ceroid lipofuscinosis
Biochem. Biophys. Res. Commun.
319
58-65
2004
Homo sapiens
Sohar, N.; Sohar, I.; Hammer, H.
Lysosomal enzyme activities: new potential markers for Sjoegrens syndrome
Clin. Biochem.
38
1120-1126
2005
Homo sapiens
Kohan, R.; Noher de Halac, I.; Anzolini, V.T.; Cismondi, A.; Oller Ramirez, A.M.; Capra, A.P.; Dodelson de Kremer, R.
Palmitoyl protein thioesterase 1 (PPT1) and tripeptidyl peptidase-I (TPP-I) are expressed in the human saliva. A reliable and non-invasive source for the diagnosis of infantile (CLN1) and late infantile (CLN2) neuronal ceroid lipofuscinoses
Clin. Biochem.
38
492-494
2005
Homo sapiens
Walus, M.; Kida, E.; Wisniewski, K.E.; Golabek, A.A.
Ser475, Glu272, Asp276, Asp327, and Asp360 are involved in catalytic activity of human tripeptidyl-peptidase I
FEBS Lett.
579
1383-1388
2005
Homo sapiens (O14773), Homo sapiens
Steinfeld, R.; Steinke, H.B.; Isbrandt, D.; Kohlschuetter, A.; Gaertner, J.
Mutations in classical late infantile neuronal ceroid lipofuscinosis disrupt transport of tripeptidyl-peptidase I to lysosomes
Hum. Mol. Genet.
13
2483-2491
2004
Homo sapiens
Oyama, H.; Fujisawa, T.; Suzuki, T.; Dunn, B.M.; Wlodawer, A.; Oda, K.
Catalytic residues and substrate specificity of recombinant human tripeptidyl peptidase I (CLN2)
J. Biochem.
138
127-134
2005
Homo sapiens
Wujek, P.; Kida, E.; Walus, M.; Wisniewski, K.E.; Golabek, A.A.
N-glycosylation is crucial for folding, trafficking, and stability of human tripeptidyl-peptidase I
J. Biol. Chem.
279
12827-12839
2004
Homo sapiens
Golabek, A.A.; Wujek, P.; Walus, M.; Bieler, S.; Soto, C.; Wisniewski, K.E.; Kida, E.
Maturation of human tripeptidyl-peptidase I in vitro
J. Biol. Chem.
279
31058-31067
2004
Homo sapiens
Golabek, A.A.; Walus, M.; Wisniewski, K.E.; Kida, E.
Glycosaminoglycans modulate activation, activity, and stability of tripeptidyl-peptidase I in vitro and in vivo
J. Biol. Chem.
280
7550-7561
2005
Homo sapiens
Tian, Y.; Sohar, I.; Taylor, J.W.; Lobel, P.
Determination of the substrate specificity of tripeptidyl-peptidase I using combinatorial peptide libraries and development of improved fluorogenic substrates
J. Biol. Chem.
281
6559-6572
2006
Homo sapiens
Steinfeld, R.; Fuhrmann, J.C.; Gaertner, J.
Detection of tripeptidyl peptidase I activity in living cells by fluorogenic substrates
J. Histochem. Cytochem.
54
991-996
2006
Homo sapiens
Kyttaelae, A.; Lahtinen, U.; Braulke, T.; Hofmann, S.L.
Functional biology of the neuronal ceroid lipofuscinoses (NCL) proteins
Biochim. Biophys. Acta
1762
920-933
2006
Homo sapiens
Golabek, A.A.; Kida, E.
Tripeptidyl-peptidase I in health and disease
Biol. Chem.
387
1091-1099
2006
Homo sapiens
Schroeder, B.; Elsaesser, H.P.; Schmidt, B.; Hasilik, A.
Characterisation of lipofuscin-like lysosomal inclusion bodies from human placenta
FEBS Lett.
581
102-108
2007
Homo sapiens (O14773), Homo sapiens
Leman, A.R.; Polochock, S.; Mole, S.E.; Pearce, D.A.; Rothberg, P.G.
Homogeneous PCR nucleobase quenching assays to detect four mutations that cause neuronal ceroid lipofuscinosis: T75P and R151X in CLN1, and IVS5-1G>C and R208X in CLN2
J. Neurosci. Methods
157
124-131
2006
Homo sapiens
Bessa, C.; Teixeira, C.A.; Dias, A.; Alves, M.; Rocha, S.; Lacerda, L.; Loureiro, L.; Guimaraes, A.; Ribeiro, M.G.
CLN2/TPP1 deficiency: the novel mutation IVS7-10A>G causes intron retention and is associated with a mild disease phenotype
Mol. Genet. Metab.
93
66-73
2008
Homo sapiens
Cabrera-Salazar, M.A.; Roskelley, E.M.; Bu, J.; Hodges, B.L.; Yew, N.; Dodge, J.C.; Shihabuddin, L.S.; Sohar, I.; Sleat, D.E.; Scheule, R.K.; Davidson, B.L.; Cheng, S.H.; Lobel, P.; Passini, M.A.
Timing of therapeutic intervention determines functional and survival outcomes in a mouse model of late infantile batten disease
Mol. Ther.
15
1782-1788
2007
Homo sapiens (O14773), Homo sapiens
Sondhi, D.; Hackett, N.R.; Peterson, D.A.; Stratton, J.; Baad, M.; Travis, K.M.; Wilson, J.M.; Crystal, R.G.
Enhanced survival of the LINCL mouse following CLN2 gene transfer using the rh.10 rhesus macaque-derived adeno-associated virus vector
Mol. Ther.
15
481-491
2007
Homo sapiens
Chang, M.; Cooper, J.D.; Sleat, D.E.; Cheng, S.H.; Dodge, J.C.; Passini, M.A.; Lobel, P.; Davidson, B.L.
Intraventricular enzyme replacement improves disease phenotypes in a mouse model of late infantile neuronal ceroid lipofuscinosis
Mol. Ther.
16
649-656
2008
Homo sapiens
Golabek, A.A.; Dolzhanskaya, N.; Walus, M.; Wisniewski, K.E.; Kida, E.
Prosegment of tripeptidyl peptidase I is a potent, slow-binding inhibitor of its cognate enzyme
J. Biol. Chem.
283
16497-16504
2008
Homo sapiens
Pal, A.; Kraetzner, R.; Gruene, T.; Grapp, M.; Schreiber, K.; Gronborg, M.; Urlaub, H.; Becker, S.; Asif, A.R.; Gaertner, J.; Sheldrick, G.M.; Steinfeld, R.
Structure of tripeptidyl-peptidase I provides insight into the molecular basis of late infantile neuronal ceroid lipofuscinosis
J. Biol. Chem.
284
3976-3984
2009
Homo sapiens (O14773)
Guhaniyogi, J.; Sohar, I.; Das, K.; Stock, A.M.; Lobel, P.
Crystal structure and autoactivation pathway of the precursor form of human tripeptidyl-peptidase 1, the enzyme deficient in late infantile ceroid lipofuscinosis
J. Biol. Chem.
284
3985-3997
2009
Homo sapiens (O14773), Homo sapiens
Walus, M.; Kida, E.; Golabek, A.A.
Functional consequences and rescue potential of pathogenic missense mutations in tripeptidyl peptidase I
Hum. Mutat.
31
710-721
2010
Homo sapiens (O14773), Homo sapiens
Lukacs, Z.; Nickel, M.; Murko, S.; Nieves Cobos, P.; Schulz, A.; Santer, R.; Kohlschuetter, A.
Validity of a rapid and simple fluorometric tripeptidyl peptidase 1 (TPP1) assay using dried blood specimens to diagnose CLN2 disease
Clin. Chim. Acta
492
69-71
2019
Homo sapiens (O14773), Homo sapiens
Kondo, M.Y.; Gouvea, I.E.; Okamoto, D.N.; Santos, J.A.; Souccar, C.; Oda, K.; Juliano, L.; Juliano, M.A.
Analysis of catalytic properties of tripeptidyl peptidase I (TTP-I), a serine carboxyl lysosomal protease, and its detection in tissue extracts using selective FRET peptide substrate
Peptides
76
80-86
2016
Homo sapiens (O14773), Rattus norvegicus (Q9EQV6), Rattus norvegicus Wistar (Q9EQV6)
Sole-Domenech, S.; Rojas, A.V.; Maisuradze, G.G.; Scheraga, H.A.; Lobel, P.; Maxfield, F.R.
Lysosomal enzyme tripeptidyl peptidase 1 destabilizes fibrillar Abeta by multiple endoproteolytic cleavages within the beta-sheet domain
Proc. Natl. Acad. Sci. USA
115
1493-1498
2018
Homo sapiens (O14773)
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