Expression and comparative analysis of recombinant rat and mouse RT6 T cell mono(ADP-ribosyl)transferases in E. coli.
Recombinant RT6 proteins of rat and mouse were analyzed for NAD-metabolizing, i.e. mono(ADP-ribosyl)transferase, NAD-glycohydrolase (NADase) and ADP-ribosyl cyclase activities. The results reveal surprising intra- as well as inter-species differences in enzyme activities. While mouse Rt6 proteins were found to be strong arginine-specific transferases, but comparatively weak NADases, the opposite held true for rat RT6, for which transferase activity could only be detected in the form of arginine-specific auto-ADP-ribosylation, displayed by RT6.2 but not by RT6.1. NADase activity of rat RT6 was not accompanied by production of cyclic ADPR (cADPR). Rat RT6 gained potent arginine-specific transferase activity by exchange of a single amino acid for the corresponding residue of the mouse proteins.
Karsten
S
S
Department of Physiological Chemistry, Hamburg University, Germany.
Schröder
J
J
da Silva
C
C
Kahlke
D
D
Thiele
H G
HG
Koch-Noite
F
F
Haag
F
F
D036002:ADP Ribose Transferases / Q000235:genetics / Q000378:metabolism*; D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000378:metabolism; D001665:Binding Sites; D004926:Escherichia coli; D015870:Gene Expression; D018698:Glutamic Acid; D006801:Humans; D008562:Membrane Glycoproteins; D051379:Mice; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D009244:NAD+ Nucleosidase / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000201:enzymology*
D015703:Antigens, CD; D000943:Antigens, Differentiation; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D018698:Glutamic Acid; D036002:ADP Ribose Transferases; D009699:N-Glycosyl Hydrolases; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; C498155:Cd38 protein, mouse; C498156:Cd38 protein, rat; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1