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 autoADP-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.
KeywordsHydrolysis HPLC Adenosine Arginine Glutamine
Unable to display preview. Download preview PDF.
- 5.Haag, F., V. Andresen, S. Karsten, F. Koch-Nolte & H.-G. Thiele. 1995. Both allelic forms of the rat T cell differentiation marker RT6 display nicotinamide adenine dinucleotide (NAD)-glycohydrolase activity, yet only RT6.2 is capable of automodification upon incubation with NAD. Eur. J. Immunol. 25: 2355.PubMedCrossRefGoogle Scholar
- 7.Koch-Nolte, F., D. Petersen, S. Balasubramanian, F. Haag, D. Kahlke, T. Wilier, R. Kastelein, F. Bazan & H.-G. Thiele. 1996. Mouse T cell membrane proteins Rt6-1 and Rt6-2 are arginine/protein mono(ADPribosyl) transferases and share secondary structure motifs with ADP-ribosylating bacterial toxins. J Biol Chem 271: 7686.PubMedCrossRefGoogle Scholar
- 9.Zocchi, E., L. Franco, L. Guida, U. Benatti, A. Bargellesi, F. Malavasi, H. C. Lee & F. A. De. 1993. A single protein immunologically identified as CD38 displays NAD+ glycohydrolase, ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities at the outer surface of human erythrocytes. Biochem Biophys Res Commun 196: 1459.PubMedCrossRefGoogle Scholar
- 19.Moss, J. & M. Vaughan. 1990. ADP-ribosylating toxins and G proteins: Insights into signal transduction. Washington DC: American Society for Microbiology.Google Scholar