Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli as GST-tagged proteins | Hathewaya limosa |
Protein Variants | Comment | Organism |
---|---|---|
E219Q | site-directed mutagenesis, inactive mutant | Hathewaya limosa |
Q217E | site-directed mutagenesis, the point mutation alters the asparagine-specificity of the enzyme to an arginie-specificity, the mutant does not show ADP-ribosyltransferase activity towards RhoA, but is still capable of NAD-binding and possesses NAD+ glycohydrolase activity, the mutant exoenzyme C3 is capable of ADP-ribosylation of poly-arginine, but not poly-asparagine, and shows high activity with Arg residues of soybean trypsin inhibitor | Hathewaya limosa |
R151A | site-directed mutagenesis, the mutation does not alter the enzyme activity or its potential as substrate for the C3 mutant Q217E | Hathewaya limosa |
R61A | site-directed mutagenesis, the mutation does not alter the enzyme activity or its potential as substrate for the C3 mutant Q217E | Hathewaya limosa |
R86A | site-directed mutagenesis, the mutation does not alter the enzyme activity but its potential as substrate for the C3 mutant Q217E, which cannot ADP-ribosylate this mutant | Hathewaya limosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Hathewaya limosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
NAD+ + (ADP-D-ribosyl)n-acceptor | Hathewaya limosa | - |
nicotinamide + (ADP-D-ribosyl)n+1-acceptor | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Hathewaya limosa | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
NAD+ + (ADP-D-ribosyl)n-acceptor = nicotinamide + (ADP-D-ribosyl)n+1-acceptor + H+ | E219 and Gln217 of the ARTT-motif, i.e. ADP-ribosylating turn-turn motif, are essential for activity | Hathewaya limosa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
80 | - |
wild-type enzyme, substrate RhoA | Hathewaya limosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme also shows NAD+-hydrolase activity | Hathewaya limosa | ? | - |
? | |
NAD+ + (ADP-D-ribosyl)n-acceptor | - |
Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)n+1-acceptor | - |
? | |
NAD+ + GTPase RhoA | the exoenzyme modifies the low-molecular-mass GTPases RhoA, B, and C specifically at Asn41 | Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)-GTPase RhoA | - |
? | |
NAD+ + GTPase RhoB | the exoenzyme modifies the low-molecular-mass GTPases RhoA, B, and C specifically at Asn41 | Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)-GTPase RhoB | - |
? | |
NAD+ + GTPase RhoC | the exoenzyme modifies the low-molecular-mass GTPases RhoA, B, and C specifically at Asn41 | Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)-GTPase RhoC | - |
? | |
NAD+ + soybean trypsin inhibitor | the exoenzyme modifies the substrate at an arginine residue | Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)-soybean trypsin inhibitor | - |
? | |
NAD+ + wild-type exoenzyme C3 | activity of the recombinant mutant Q217E exoenzyme C3, modification at Arg86 | Hathewaya limosa | nicotinamide + (ADP-D-ribosyl)-wild-type exoenzyme C3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
asparagine-specific ADP-ribosyltransferase | - |
Hathewaya limosa |
C3lim | - |
Hathewaya limosa |
exoenzyme C3 | - |
Hathewaya limosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 37 | assay at | Hathewaya limosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Hathewaya limosa |