Q8GB88 · SCPA_ALISE
- ProteinKumamolisin-As
- GenescpA
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids553 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Thermostable pepstatin-insensitive serine-carboxyl proteinase (PubMed:12513991, PubMed:15014068).
Can specifically and efficiently degrade collagen (PubMed:12513991, PubMed:15014068).
In vitro, preferentially cleaves the bovine collagen alpha-1 chain at a single specific cleavage site (PubMed:15014068).
Can also hydrolyze the chromogenic substrate azocoll and synthetic peptides derived from collagen (PubMed:12513991, PubMed:15014068).
Also shows low activity with albumin and casein, but cannot use keratin azure and elastin (PubMed:12513991).
May play a saprophytic role in soils through degradation of collagenous materials (such as those from carcasses) in cooperation with other saprophytes under thermoacidophilic conditions and play an important role in global cycling of nitrogen (PubMed:15014068).
Can specifically and efficiently degrade collagen (PubMed:12513991, PubMed:15014068).
In vitro, preferentially cleaves the bovine collagen alpha-1 chain at a single specific cleavage site (PubMed:15014068).
Can also hydrolyze the chromogenic substrate azocoll and synthetic peptides derived from collagen (PubMed:12513991, PubMed:15014068).
Also shows low activity with albumin and casein, but cannot use keratin azure and elastin (PubMed:12513991).
May play a saprophytic role in soils through degradation of collagenous materials (such as those from carcasses) in cooperation with other saprophytes under thermoacidophilic conditions and play an important role in global cycling of nitrogen (PubMed:15014068).
Miscellaneous
Kumamolisin-As does not make a specific three-quarters/one-quarter cut of the triple-helical, native collagen, characteristic for the previously described collagenases.
Cofactor
Note: Binds 1 Ca2+ ion per subunit.
Activity regulation
Partially inhibited by L-alanyl-L-alanyl-L-phenylalanyl-chloromethane, leupeptin, 1 mM Fe2+ or 1 mM Hg2+ (PubMed:12513991).
Insensitive to specific inhibitors of aspartic proteinases, such as pepstatin and diazoacetyl-DL-norleucine methyl ester (DAN), to specific inhibitors of metalloproteinases (EDTA), cysteine proteinases (N-ethylmaleimide) and serine proteinases (PMSF) (PubMed:12513991).
Insensitive to specific inhibitors of aspartic proteinases, such as pepstatin and diazoacetyl-DL-norleucine methyl ester (DAN), to specific inhibitors of metalloproteinases (EDTA), cysteine proteinases (N-ethylmaleimide) and serine proteinases (PMSF) (PubMed:12513991).
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
32 μM | Phe-Gly-Pro-Ala-Gly-Pro-Ile-Gly | |||||
214 μM | Met-Gly-Pro-Arg-Gly-Phe-Pro-Gly-Ser | |||||
1 μM | (N-methylanthranilic acid)-Met-Gly-Pro-His-Phe-Phe-Pro-N-(2,4-dinitrophenyl)-Lys-D-Arg-D-Arg (IQF) | 4.0 | 40 |
kcat is 5.41 sec-1 with Phe-Gly-Pro-Ala-Gly-Pro-Ile-Gly as substrate (PubMed:12513991).
kcat is 351 sec-1 with Met-Gly-Pro-Arg-Gly-Phe-Pro-Gly-Ser as substrate (PubMed:12513991).
kcat is 395 sec-1 with IQF as substrate (at 40 degrees Celsius and pH 4.0) (PubMed:15014068).
kcat is 351 sec-1 with Met-Gly-Pro-Arg-Gly-Phe-Pro-Gly-Ser as substrate (PubMed:12513991).
kcat is 395 sec-1 with IQF as substrate (at 40 degrees Celsius and pH 4.0) (PubMed:15014068).
pH Dependence
Optimum pH is 3.9 (for degradation of azocoll) (PubMed:12513991).
Stable at pH 3.5 to 5.0 (PubMed:12513991).
Stable at pH 3.5 to 5.0 (PubMed:12513991).
Temperature Dependence
Optimum temperature is 60 degrees Celsius (for degradation of azocoll).
Features
Showing features for active site, binding site.
Type | ID | Position(s) | Description | ||
---|---|---|---|---|---|
Active site | 267 | Charge relay system | |||
Active site | 271 | Charge relay system | |||
Active site | 467 | Charge relay system | |||
Binding site | 505 | Ca2+ (UniProtKB | ChEBI) | |||
Binding site | 506 | Ca2+ (UniProtKB | ChEBI) | |||
Binding site | 523 | Ca2+ (UniProtKB | ChEBI) | |||
Binding site | 525 | Ca2+ (UniProtKB | ChEBI) | |||
Binding site | 527 | Ca2+ (UniProtKB | ChEBI) | |||
GO annotations
Aspect | Term | |
---|---|---|
Molecular Function | metal ion binding | |
Molecular Function | serine-type endopeptidase activity | |
Molecular Function | tripeptidyl-peptidase activity | |
Biological Process | proteolysis |
Keywords
- Molecular function
- Ligand
Enzyme and pathway databases
Protein family/group databases
Names & Taxonomy
Protein names
- Recommended nameKumamolisin-As
- EC number
- Alternative names
Gene names
Organism names
- Organism
- Strain
- Taxonomic lineageBacteria > Bacillota > Bacilli > Bacillales > Alicyclobacillaceae > Alicyclobacillus
Accessions
- Primary accessionQ8GB88
Subcellular Location
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | ||
---|---|---|---|---|---|
Mutagenesis | 267 | Loses both auto-processing activity and proteolytic activity; when associated with N-353. | |||
Mutagenesis | 267 | Can be very slowly converted during the acid treatment into a mature form. Almost loss of activity with IQF as substrate. Exists as an equilibrated mixture of nicked and intact forms of the precursor and retains very low activity; when associated with N-353. | |||
Mutagenesis | 353 | Retains auto-processing activity but only low activity. Exists as an equilibrated mixture of nicked and intact forms of the precursor and retains very low activity; when associated with H-267. Loses both auto-processing activity and proteolytic activity; when associated with A-267 or Q-267. | |||
PTM/Processing
Features
Showing features for propeptide, chain.
Type | ID | Position(s) | Description | ||
---|---|---|---|---|---|
Propeptide | PRO_0000461157 | 1-189 | Removed in mature form | ||
Chain | PRO_0000461158 | 190-553 | Kumamolisin-As | ||
Post-translational modification
Autocatalytically processed.
Keywords
- PTM
Expression
Induction
Constitutively expressed.
Interaction
Subunit
Monomer.
Structure
Sequence
- Sequence statusComplete
- Sequence processingThe displayed sequence is further processed into a mature form.
- Length553
- Mass (Da)57,154
- Last updated2003-03-01 v1
- MD5 Checksum64CA434B76934708EED800B6934324C9
Mass Spectrometry
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
AB085855 EMBL· GenBank· DDBJ | BAC41257.1 EMBL· GenBank· DDBJ | Genomic DNA |