Three conserved consensus sequences identify the NAD-binding site of ADP-ribosylating enzymes, expressed by eukaryotes, bacteria and T-even bacteriophages

• Published in: Molecular microbiology Vol. 21; no. 4; p. 667
• Main Authors: Domenighini, M, Rappuoli, R
• Format: Journal Article
• Language: English
• Published: England 01.08.1996
• Subjects: ADP Ribose Transferases - chemistry; Amino Acid Sequence; Binding Sites; Conserved Sequence; Molecular Sequence Data; NAD - metabolism; Poly(ADP-ribose) Polymerases - chemistry
• ISSN: 0950-382X
• DOI: 10.1046/j.1365-2958.1996.321396.x

It has been previously reported that the three-dimensional structures of the NAD-binding and catalytic site of bacterial toxins with ADP-ribosylating activity are superimposable, and that the key amino acids for the enzymatic activity are conserved. The model includes an NAD-binding and catalytic site formed by an alpha-helix bent over a beta-strand, surrounded by two beta-strands bearing a Glu and a His, or Arg, that are required for catalysis. We show here that the model can be extended to comprise all proteins with ADP-ribosylating activity known to date, including all eukaryotic mono- and poly-ADP-ribosyltransferases, the bacterial ADP-ribosylating enzymes which do not have toxic activity, and the analogous enzymes encoded by T-even bacteriophages. We show that, in addition to the common Glu and Arg/His amino acids previously identified, the conserved motifs can be extended as follows: (i) the Arg/His motif is usually arom-His/Arg (where 'arom' is an aromatic residue); (ii) in the sequences of the CT group the beta-strand forming part of the 'scaffold' of the catalytic cavity has an arom-ph-Ser-Thr-Ser-ph consensus (where 'ph' represents a hydrophobic residue); and (iii) the motif centered in the key glutamic residue is Glu/Gin-X-Glu; while (iv) in the sequences of the DT group the NAD-binding motif is Tyr-X10-Tyr. We believe that the model proposed not only accounts for all ADP-ribosylating proteins known to date, but it is likely to fit other enzymes (currently being analysed) which possess such an activity.