Partial purification and characterization of ADP-ribosyltransferase produced by Legionella pneumophila.
A scheme for the partial purification of a Legionella pneumophila product possessing ADP-ribosyl-transferase and NAD-glycohydrolase activities is presented. The purification steps consisted of gel chromatography, ion-exchange, hydrophobic interaction chromatography, and chromatofocusing. The partially purified preparation modified eukaryotic components of molecular mass 20-25 kDa, which it is proposed are GTP-binding proteins. Addition of bivalent cations as well as ATP to the reaction buffer was necessary for ADP-ribosylation. NAD (50 microM) and nicotinamide (16 mM) greatly inhibited incorporation of ADP-ribose into acceptor proteins.
Belyi YuF
Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow.
Tartakovskii
I S
IS
Vertiev YuV
Prosorovskii
S V
SV
D001345:Autoradiography; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D007875:Legionella / Q000201:enzymology* / Q000254:growth & development; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009244:NAD+ Nucleosidase / Q000302:isolation & purification / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D011065:Poly(ADP-ribose) Polymerases / Q000302:isolation & purification* / Q000378:metabolism
D010761:Phosphorus Radioisotopes; D009243:NAD; D011065:Poly(ADP-ribose) Polymerases; D009244:NAD+ Nucleosidase