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Literature summary for 2.4.2.30 extracted from

  • Vogelsgesang, M.; Aktories, K.
    Exchange of glutamine-217 to glutamate of Clostridium limosum exoenzyme C3 turns the asparagine-specific ADP-ribosyltransferase into an arginine-modifying enzyme (2006), Biochemistry, 45, 1017-1025.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli as GST-tagged proteins Hathewaya limosa

Protein Variants

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

Metals/Ions Comment Organism Structure
Mg2+ required Hathewaya limosa

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Hathewaya limosa
-
-
-

Reaction

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 [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
80
-
wild-type enzyme, substrate RhoA Hathewaya limosa

Substrates and Products (Substrate)

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

Synonyms Comment Organism
asparagine-specific ADP-ribosyltransferase
-
Hathewaya limosa
C3lim
-
Hathewaya limosa
exoenzyme C3
-
Hathewaya limosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 37 assay at Hathewaya limosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Hathewaya limosa