Literature summary extracted from
Hao, Y.; Purtha, W.; Cortesio, C.; Rui, H.; Gu, Y.; Chen, H.; Sickmier, E.A.; Manzanillo, P.; Huang, X.
Crystal structures of human procathepsin H (2018), PLoS ONE, 13, e0200374 .
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
3.4.22.16 |
additional information |
procathepsin H is not capable of autoactivation and requires other proteases such as cathepsin L for its activation |
Homo sapiens |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.4.22.16 |
procathepsin H (Ala1P-Val220) construct is cloned with an N-terminal gp64 secretion signal peptide and an uncleavable C-terminal His6-tag, recombinant expression of wild-type enzyme in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system, recombinant expression of His-tagged mutant enzymes in HEK-293T cells using the lentiviral viral vector CD731B |
Homo sapiens |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
3.4.22.16 |
purified wild-type and C26S mutant procathepsin H, sitting drop vapor diffusion method, 5.7 mg/ml protein, crystal growth from 0.1 M Na2HPO4/citric acid, pH 4.2, and 2.0 M (NH4)2SO4, 1 week, 20°C, method optimization, X-ray diffraction structure determination and analysis at 2.0 A and 1.66 A resolution, respectively, molecular replacement and modeling |
Homo sapiens |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
3.4.22.16 |
C26S |
site-directed mutagenesis, catalytically inactive active site mutant |
Homo sapiens |
3.4.22.16 |
C80S |
site-directed mutagenesis, the mutation removes the disulfide between the mini-chain and the mature domain and completely abolishes the protease activity |
Homo sapiens |
3.4.22.16 |
H166A |
site-directed mutagenesis, catalytically inactive active site mutant |
Homo sapiens |
3.4.22.16 |
N186A |
site-directed mutagenesis, catalytically inactive active site mutant |
Homo sapiens |
3.4.22.16 |
T83A |
site-directed mutagenesis, mutation of the Thr83 of the mini-chain has only mild effects on the enzyme activity |
Homo sapiens |
General Stability
EC Number |
General Stability |
Organism |
---|
3.4.22.16 |
analysis of stability of the mini-chain in cathepsin H and procathepsin H, overview |
Homo sapiens |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.4.22.16 |
E64d |
- |
Homo sapiens |
|
3.4.22.16 |
additional information |
molecular basis for the inhibition of the mature enzyme by the prodomain. The prodomain of procathepsin H has an N-terminal helical subdomain (Ala1P-Ser75P) and an extended portion (Glu76P-Pro93P) that links the helical subdomain to Tyr1 of the mature domain |
Homo sapiens |
|
Localization
EC Number |
Localization |
Comment |
Organism |
GeneOntology No. |
Textmining |
---|
3.4.22.16 |
lysosome |
- |
Homo sapiens |
5764 |
- |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.4.22.16 |
Homo sapiens |
P09668 |
- |
- |
Posttranslational Modification
EC Number |
Posttranslational Modification |
Comment |
Organism |
---|
3.4.22.16 |
glycoprotein |
the enzyme harbors two glycosylation sites, Asn79P on the mini-chain of the prodomain and Asn115 on the mature domain. No glycan is built for Asn79P in cathepsin H due to poor electron density, although it is glycosylated |
Homo sapiens |
3.4.22.16 |
proteolytic modification |
the enzyme is synthesized as an inactive proenzyme, the N-terminal prodomain (Ala1P-Pro93P) is cleaved off for maturation, a C-terminal mature domain (Tyr1-Val220) with a disulfide bridge between Cys212 and the mini-chain Cys80P remains. Procathepsin H is not autoactivated but can be transactivated by cathepsin L. The prodomain of procathepsin H has an N-terminal helical subdomain (Ala1P-Ser75P) and an extended portion (Glu76P-Pro93P) that links the helical subdomain to Tyr1 of the mature domain |
Homo sapiens |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.4.22.16 |
recombinant His-tagged wild-type and mutant enzymes from Spodoptera frugiperda Sf9 cells or HEK-23T cells, respectively, by nickel affinity chromatography and gel filtration |
Homo sapiens |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.4.22.16 |
Arg-7-amido-4-trifluoromethylcoumarin + H2O |
fluorogenic cathepsin H substrate |
Homo sapiens |
Arg + 7-amino-4-trifluoromethylcoumarin |
- |
? |
|
3.4.22.16 |
additional information |
procathepsin H is not capable of autocleavage at pH 4.5-6.5 |
Homo sapiens |
? |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
3.4.22.16 |
procathepsin H |
- |
Homo sapiens |
General Information
EC Number |
General Information |
Comment |
Organism |
---|
3.4.22.16 |
evolution |
cathepsin H is a member of the papain superfamily of lysosomal cysteine proteases |
Homo sapiens |
3.4.22.16 |
additional information |
the catalytic triad residues Cys26, His166 and Asn186 of procathepsin H reside in the active site cleft |
Homo sapiens |
3.4.22.16 |
physiological function |
the mature enzyme is inhibited by its own prodomain after maturation cleavage. Upon activation, cathepsin H displays a distinct proteolytic activity among the papain superfamily, functioning predominantly as an aminopeptidase that cleaves a single N-terminal residue from a polypeptide chain. Cathepsin H has also been shown to act as an endopeptidase, although with much lower efficiency |
Homo sapiens |