Characterization of NAD:arginine ADP-ribosyltransferases

Abstract

NAD:arginine mono-ADP-ribosyltransferases catalyze the transfer of ADP-ribose from NAD to the guanidino group of arginine on a target protein. Deduced amino acid sequences of one family (ART1) of mammalian ADP-ribosyltransferases, cloned from muscle and lymphocytes, show hydrophobic amino and carboxyl termini consistent with glycosylphosphatidylinositol (GPI)-anchored proteins. The proteins, overexpressed in mammalian cells transfected with the transferase cDNAs, are released from the cell surface with phosphatidylinositol-specific phospholipase C (PI-PLC), and display immunological and biochemical characteristics consistent with a cell surface, GPI-anchored protein. In contrast, the deduced amino acid sequence of a second family (ART5) of transferases, cloned from murine lymphoma cells and expressed in high abundance in testis, displays a hydrophobic amino terminus, consistent with a signal sequence, but lacks a hydrophobic signal sequence at its carboxyl terminus, suggesting that the protein is destined for export. Consistent with the surface localization of the GPI-linked transferases, multiple surface substrates have been identified in myotubes and activated lymphocytes, and, notably, include integrin α subunits. Similar to the bacterial toxin ADP-ribosyltransferases, the mammalian transferases contain the characteristic domains involved in NAD binding and ADP-ribose transfer, including a highly acidic region near the carboxy terminus, which, when disrupted by in vitro mutagenesis, results in a loss of enzymatic activity. The carboxyl half of the protein, synthesized as a fusion protein in E. coli, possessed NADase, but not ADP-ribosyltransferase activity. These findings are consistent with the existence at the carboxyl terminus of ART1 of a catalytically active domain, capable of hydrolyzing NAD, but not of transferring ADP-ribose to a guanidino acceptor.

Glycosylphosphatidylinositol Anchor Modification Machinery Deficiency Is Responsible for the Formation of Pro-Prion Protein (PrP) in BxPC-3 Protein and Increases Cancer Cell Motility

“…We treated AsPC-1 and BxPC-3 cells with PI-PLC. PI-PLC treatment is a well established assay to determine whether a protein is GPI-anchored or not (11,41). Using mAb 4H2, we found that although the PrP on the cell surface of BxPC-3 was highly resistant to PI-PLC, the PrP on the surface of AsPC-1 was much more susceptible to identical treatment ( Fig.…”

PARP12-catalyzed mono-ADP-ribosylation of Golgin-97 controls the transport of E-cadherin

“…MARylation was identified decades ago as a protein, post-translational modification (PTM) exploited by bacterial toxins to promote bacterial pathogenesis in host cells (Gill et al, 1969;Moss and Vaughan, 1977). While the occurrence of this protein PTM in mammalian cells has soon become evident (Dani et al, 2011;Jones and Baird, 1997;Lupi et al, 2000;Lupi et al, 2002;Moss et al, 1999;Okazaki and Moss, 1996;Seman et al, 2004) only recently, with the definition of the different PARPs (and of their intrinsic enzymatic activities) the physiological functions the MARylating PARPs may regulate are emerging (Cohen and Chang, 2018;Gupte et al, 2017).…”

Expression of ADP-ribosyltransferase 1 Is Associated with Poor Prognosis of Glioma Patients

“…Mono-ADP-ribosylation is a type of protein posttranslational modification (PTM), which can trans fer a single ADP-ribose from nicotin amide adenine dinucleotide (NAD + ) to the specific amino-acids residues of target proteins (Zolkiewska et al 1992;Moss et al 1999). PTM plays an important role in various physiological and pathophysiological processes, such as DNA repair, transcription, cell cycle regulation, signal transduc tion, necrosis and apoptosis (Hassa et al 2006).…”