Molecular and Immunological Characterization of ADP-ribosylarginine Hydrolases

Moss, Joel; Stanley, S J; Nightingale, Maria S.; Murtagh, James J.; Monaco, Luca Proietti; Mishima, Kōichi; Chen, Hao-Chia; Williamson, Kristin C.; Tsai, Su-Chen

doi:10.1007/978-1-4419-8718-1_70

Cited by 65 publications

(79 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…8 and 9), in accord with previous studies (56,71). Enzymes capable of reversing ADP-ribosylation include ADP-ribosylhydrolases which can remove the entire ADP-ribose moiety (72,73) and phosphodiesterases which remove only AMP, leaving ribose phosphate attached to the target protein (74). To date ADP-ribosylhydrolases have been described only as intracellular proteins (75), whereas phosphodiesterase isoforms have been cloned and characterized that function as membrane bound and secretory ecto-enzymes (76,77).…”

Section: Discussionsupporting

confidence: 79%

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

Scheuplein

Schwarz²,

Adriouch

et al. 2009

The Journal of Immunology

115

136

View full text Add to dashboard Cite

Extracellular NAD+ and ATP trigger the shedding of CD62L and the externalization of phosphatidylserine on murine T cells. These events depend on the P2X7 ion channel. Although ATP acts as a soluble ligand to activate P2X7, gating of P2X7 by NAD+ requires ecto-ADP-ribosyltransferase ART2.2-catalyzed transfer of the ADP-ribose moiety from NAD+ onto Arg125 of P2X7. Steady-state concentrations of NAD+ and ATP in extracellular compartments are highly regulated and usually are well below the threshold required for activating P2X7. The goal of this study was to identify possible endogenous sources of these nucleotides. We show that lysis of erythrocytes releases sufficient levels of NAD+ and ATP to induce activation of P2X7. Dilution of erythrocyte lysates or incubation of lysates at 37°C revealed that signaling by ATP fades more rapidly than that by NAD+. We further show that the routine preparation of primary lymph node and spleen cells induces the release of NAD+ in sufficient concentrations for ART2.2 to ADP-ribosylate P2X7, even at 4°C. Gating of P2X7 occurs when T cells are returned to 37°C, rapidly inducing CD62L-shedding and PS-externalization by a substantial fraction of the cells. The “spontaneous” activation of P2X7 during preparation of primary T cells could be prevented by i.v. injection of either the surrogate ART substrate etheno-NAD or ART2.2-inhibitory single domain Abs 10 min before sacrificing mice.

show abstract

Section: Discussionsupporting

confidence: 79%

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

Scheuplein

Schwarz²,

Adriouch

et al. 2009

The Journal of Immunology

115

136

View full text Add to dashboard Cite

show abstract

“…The two partial amino acid sequences of NAD+-glycohydrolase which were obtained after proteolytic digestion of the protein band and subsequent gas phase sequencing showed high similarities to two sequence stretches of another mammalian ADPR metabolizing enzyme, namely an arginine ADPR hydrolase from rat brain [21]. In our view, this finding corroborates our assignment of the enzyme to the protein with a molar mass of 32,000.…”

Section: Discussionsupporting

confidence: 86%

“…For a more detailed database search we used the program FINDPATTERNS of the GCG program package (Genetics Computer Group, 575 Science Drive, Madison, WI 53711, USA) to screen for proteins containing sequences similar to both peptides with the restriction that such a double hit in one protein should occur within 300 amino acids, a length which corresponds to the putative length of the amino acid sequence of the purified protein with a molar mass of 32,000. Under these conditions the top scoring protein in all databases was a murine arginine ADPR hydrolase [21]. This protein has a molar mass of 39,000 and harbours 2 sequences which are highly similar to the sequences determined for the mitochondrial NAD+-glycohydrolase and occur within a distance of 55 amino acids (Fig.…”

Section: Resultsmentioning

confidence: 98%

See 1 more Smart Citation

Identification and purification of a bovine liver mitochondrial NAD⁺‐glycohydrolase

et al. 1995

View full text Add to dashboard Cite

Nonenzymatic ADP-ribosylation of mitochondrial proteins is thought to play a role in the regulation of Ca 2÷ efflux from mitochondria. It has been shown that intramitochondrial ADP-ribose is generated by a specific NAD+glycohydrolase, which catalizes hydrolysis of NAD ÷ to ADP-ribose and nicotinamide. We purified this enzyme from bovine liver mitocbondrial membranes. The final preparation had a 1660-fold purified enzyme activity and contained a main protein band with an apparent molar mass of 32,000 in a SDS-polyacrylamide gel. The identity of this protein band with NAD+-glycohydrolase was verified by renaturation of its enzymatic activity. Partial amino acid sequence information was obtained from two enzyme fragments after proteolytic cleavage of the protein band in the SDS-polyacrylamide gel. Searches in protein databases revealed that an arginine ADP-ribosyl hydrolase harbours two stretches of amino acids that are highly similar to the partial NAD÷-glycohydrolase sequences.

show abstract

“…The human genome encodes three DRAG-related proteins designated ARH1, ARH2, and ARH3 (19), which are 357, 354, and 363 residues long, respectively. ARH1, like DRAG, specifically de-ADP-ribosylates proteins mono-ADP-ribosylated on arginine residues (20,21). ARH3 de-ADP-ribosylates polyADP-ribosylated proteins, albeit at only Ϸ10% of the activity observed for the PARG (21), with which it shares little if any sequence similarity (19,21).…”

mentioning

confidence: 99%

The structure of human ADP-ribosylhydrolase 3 (ARH3) provides insights into the reversibility of protein ADP-ribosylation

Mueller-Dieckmann

Kernstock²,

Lisurek

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

100

136

View full text Add to dashboard Cite

Posttranslational modifications are used by cells from all kingdoms of life to control enzymatic activity and to regulate protein function. For many cellular processes, including DNA repair, spindle function, and apoptosis, reversible mono-and polyADP-ribosylation constitutes a very important regulatory mechanism. Moreover, many pathogenic bacteria secrete toxins which ADP-ribosylate human proteins, causing diseases such as whooping cough, cholera, and diphtheria. Whereas the 3D structures of numerous ADP-ribosylating toxins and related mammalian enzymes have been elucidated, virtually nothing is known about the structure of protein de-ADP-ribosylating enzymes. Here, we report the 3D structure of human ADP-ribosylhydrolase 3 (hARH3). The molecular architecture of hARH3 constitutes the archetype of an all-␣-helical protein fold and provides insights into the reversibility of protein ADP-ribosylation. Two magnesium ions flanked by highly conserved amino acids pinpoint the active-site crevice. Recombinant hARH3 binds free ADP-ribose with micromolar affinity and efficiently de-ADP-ribosylates poly-but not monoADP-ribosylated proteins. Docking experiments indicate a possible binding mode for ADP-ribose polymers and suggest a reaction mechanism. Our results underscore the importance of endogenous ADP-ribosylation cycles and provide a basis for structure-based design of ADP-ribosylhydrolase inhibitors.protein structure ͉ posttranslational modification ͉ glycohydrolase ͉ docking P osttranslational modifications (PTMs) are covalent modifications of amino acid side chains in proteins that come in many different sizes and shapes, ranging from the simple addition of a phosphate group to complex multistep glycosylations. Enzyme-catalyzed PTMs allow rapid responses to environmental stimuli and play crucial roles in signal transduction. NAD-dependent ADP-ribosylation is a reversible PTM in which mono-and polyADP-ribosyltransferases (ARTs and PARPs) and ADP-ribosylhydrolases (ARHs) and poly(ADP-ribose) glycohydrolases (PARGs) catalyze amino acid-specific ADPribosylation and de-ADP-ribosylation, respectively ( Fig. 1) (1-10). ADP-ribosylation has attracted attention because bacterial virulence factors, including diphtheria, cholera, and pertussis toxin, use it as part of their pathogenic mechanism (2, 11). Mono-and polyADP-ribosylation have been recognized also as regulatory mechanisms in many cellular processes, including DNA-repair, chromatin decondensation, transcription, telomere function, mitotic spindle formation, and apoptosis (5-10, 12).Several enzymes have been cloned that catalyze de-ADPribosylation of mono-or polyADP-ribosylated proteins (13-15). Dinitrogenase-activating glycohydrolase (DRAG), an Arg-specific ARH from the phototrophic bacterium Rhodospirillum rubrum, regulates a key enzyme of nitrogen fixation (16-18). The human genome encodes three DRAG-related proteins designated ARH1, ARH2, and ARH3 (19), which are 357, 354, and 363 residues long, respectively. ARH1, like DRAG, specifically de-ADP-ribosylates prote...

show abstract

Molecular and Immunological Characterization of ADP-ribosylarginine Hydrolases

Cited by 65 publications

References 10 publications

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

Identification and purification of a bovine liver mitochondrial NAD⁺‐glycohydrolase

The structure of human ADP-ribosylhydrolase 3 (ARH3) provides insights into the reversibility of protein ADP-ribosylation

Contact Info

Product

Resources

About

Molecular and Immunological Characterization of ADP-ribosylarginine Hydrolases

Cited by 65 publications

References 10 publications

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

NAD+ and ATP Released from Injured Cells Induce P2X7-Dependent Shedding of CD62L and Externalization of Phosphatidylserine by Murine T Cells

Identification and purification of a bovine liver mitochondrial NAD+‐glycohydrolase

The structure of human ADP-ribosylhydrolase 3 (ARH3) provides insights into the reversibility of protein ADP-ribosylation

Contact Info

Product

Resources

About

Identification and purification of a bovine liver mitochondrial NAD⁺‐glycohydrolase