Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | deglycosylation enhances CmCatB activity, but compromises CmCatB stability | Callosobruchus maculatus |
Cloned (Comment) | Organism |
---|---|
functional expression of His-tagged wild-type CatB in Pichia pastoris, and expression of some His-tagged glycosylation site mutants. Residue Asn236 appears necessary for protein expression | Callosobruchus maculatus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | simultaneously replacing all three Asn residue N-glycosylation sites with Gln via site-directed mutagenesis does not result in completely unglycosylated protein due to the existence of additional atypical glycosylation sites | Callosobruchus maculatus |
N100Q | site-directed mutagenesis, the mutation substantially enhances proteolytic activity but compromises protein stability | Callosobruchus maculatus |
N111Q | site-directed mutagenesis | Callosobruchus maculatus |
N207Q | site-directed mutagenesis | Callosobruchus maculatus |
N236Q | site-directed mutagenesis | Callosobruchus maculatus |
N97Q | site-directed mutagenesis | Callosobruchus maculatus |
N97Q/N100Q/N207Q | site-directed mutagenesis | Callosobruchus maculatus |
N97Q/N100Q/N207Q/N236Q | site-directed mutagenesis, the mutant cannot be expressed in Pichia pastoris cells | Callosobruchus maculatus |
N97Q/N100Q/N236Q | site-directed mutagenesis | Callosobruchus maculatus |
N97Q/N207Q | site-directed mutagenesis | Callosobruchus maculatus |
N97Q/N207Q/N236Q | site-directed mutagenesis, the mutant cannot be expressed in Pichia pastoris cells | Callosobruchus maculatus |
General Stability | Organism |
---|---|
deglycosylation compromises CmCatB stability | Callosobruchus maculatus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Callosobruchus maculatus | - |
cowpea bruchid, a grain storage pest | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | glycosylation at atypical Asn-X-Cys motif, non-canonical Asn-X-Cys sites, wild-type CatB contains three N-glycosylation Asn-X-Ser/Thr consensus sequences with existence of additional atypical glycosylation sites. The heterogeneously expressed CazB is also glycosylated in Pichia patoris. Residue Asn100 is responsible for non-canonical glycosylation. Glycosylation at the non-canonical Asn100 site decreases the proteolytic activity of CmCatB, while it enhances protein stability. Deglycosylation enhances CmCatB activity, but compromises CmCatB stability | Callosobruchus maculatus |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant CatBs from Pichia pastoris by nickel affinity chromatography and ammonium sulfate fractionation | Callosobruchus maculatus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
Hydrolysis of proteins with broad specificity for peptide bonds. Preferentially cleaves -Arg-Arg-/- bonds in small molecule substrates (thus differing from cathepsin L). In addition to being an endopeptidase, shows peptidyl-dipeptidase activity, liberating C-terminal dipeptides | the catalytic residue is Cys29 | Callosobruchus maculatus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
protein rGSII-D88N + H2O | degradation | Callosobruchus maculatus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | tertiary structure model of CmCatB., overview | Callosobruchus maculatus |
Synonyms | Comment | Organism |
---|---|---|
CatB | - |
Callosobruchus maculatus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Callosobruchus maculatus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
recombinant enzyme | Callosobruchus maculatus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4.5 | 5.5 | maxinal activity at pH 4.5, inactivation at pH 5.5 and inactive above | Callosobruchus maculatus |
General Information | Comment | Organism |
---|---|---|
physiological function | the cysteine protease CatB facilitates insects coping with dietary protease inhibitor challenge | Callosobruchus maculatus |