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Literature summary for 3.4.22.1 extracted from

  • Chi, Y.H.; Koo, Y.D.; Dai, S.Y.; Ahn, J.E.; Yun, D.J.; Lee, S.Y.; Zhu-Salzman, K.
    N-glycosylation at non-canonical Asn-X-Cys sequence of an insect recombinant cathepsin B-like counter-defense protein (2010), Comp. Biochem. Physiol. B, 156, 40-47.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information deglycosylation enhances CmCatB activity, but compromises CmCatB stability Callosobruchus maculatus

Cloned(Commentary)

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

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

General Stability Organism
deglycosylation compromises CmCatB stability Callosobruchus maculatus

Organism

Organism UniProt Comment Textmining
Callosobruchus maculatus
-
cowpea bruchid, a grain storage pest
-

Posttranslational Modification

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 (Commentary)

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

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
protein rGSII-D88N + H2O degradation Callosobruchus maculatus ?
-
?

Subunits

Subunits Comment Organism
More tertiary structure model of CmCatB., overview Callosobruchus maculatus

Synonyms

Synonyms Comment Organism
CatB
-
Callosobruchus maculatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Callosobruchus maculatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.5
-
recombinant enzyme Callosobruchus maculatus

pH Range

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

General Information Comment Organism
physiological function the cysteine protease CatB facilitates insects coping with dietary protease inhibitor challenge Callosobruchus maculatus