Publications

• Reversibility of arginine-specific mono(ADP-ribosyl)ation: identification in erythrocytes of an ADP-ribose-L-arginine cleavage enzyme.

  Moss J., Jacobson M.K., Stanley S.J.

  Enzymes have been identified in animal tissues that catalyze the mono(ADP-ribosyl)ation of arginine and proteins. Since these NAD:arginine ADP-ribosyltransferases under physiological conditions do not appear to catalyze the degradation of the product ADP-ribose-arginine, the possibility was invest ... >> More

  Enzymes have been identified in animal tissues that catalyze the mono(ADP-ribosyl)ation of arginine and proteins. Since these NAD:arginine ADP-ribosyltransferases under physiological conditions do not appear to catalyze the degradation of the product ADP-ribose-arginine, the possibility was investigated that a different family of enzymes exists that cleaves the ADP-ribose-arginine linkage. An enzyme was identified in and partially purified from turkey erythrocytes that catalyzed the degradation of ADP-ribose-[14C]arginine synthesized by a salt-activated NAD:arginine ADP-ribosyl-transferase, resulting in the release of a radiolabeled compound that was characterized chromatographically and by amino acid analysis as arginine. This putative arginine product was converted in a reaction dependent on NAD and the NAD:arginine ADP-ribosyltransferase to a compound exhibiting properties characteristic of ADP-ribose-arginine. Action of cleavage enzyme on [adenine-U-14C]ADP-ribose-arginine resulted in the release of a radiolabeled compound that behaved chromatographically like [adenine-U-14C]ADP-ribose. Since degradation of ADP-ribose-arginine appears to generate an arginine moiety that is a substrate for the NAD:arginine ADP-ribosyltransferase, it appears that ADP-ribosylation may be a reversible modification of proteins. << Less

  Proc Natl Acad Sci U S A 82:5603-5607(1985) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Detection of arginine-ADP-ribosylated protein using recombinant ADP-ribosylarginine hydrolase.

  Ohno T., Tsuchiya M., Osago H., Hara N., Jidoi J., Shimoyama M.

  We made use of ADP-ribosylarginine hydrolase to detect arginine-ADP-ribosylated proteins. The hydrolase was expressed in Escherichia coli as a protein fused with glutathione S-transferase (GST). The fusion protein GST-ADP-ribosylarginine hydrolase catalyzed the hydrolysis of alpha-ADP-ribosylargin ... >> More

  We made use of ADP-ribosylarginine hydrolase to detect arginine-ADP-ribosylated proteins. The hydrolase was expressed in Escherichia coli as a protein fused with glutathione S-transferase (GST). The fusion protein GST-ADP-ribosylarginine hydrolase catalyzed the hydrolysis of alpha-ADP-ribosylarginine to produce ADP-ribose and arginine. Casein ADP-ribosylated with [32P]NAD and chicken heterophil arginine-specific ADP-ribosyltransferase served as a substrate for the recombinant ADP-ribosylarginine hydrolase and the released ADP-ribose was determined. Protein ADP-ribosylated by cholera toxin could serve as substrate of the hydrolase but protein ADP-ribosylated by pertussis toxin, diphtheria toxin, or C(3) enzyme of Clostridium botulinum could not. The hydrolase did not release the radioactivity incorporated into isolated rat liver nuclei incubated with [(32)P]NAD or in bovine brain cytosol incubated with [(32)P]ADP-ribose. In homogenate of mouse heart which contained arginine-specific ADP-ribosyltransferase, labeling of a 55-kDa protein by incubation with [(32)P]NAD was removed by ADP-ribosylarginine hydrolase treatment; hence, the specific hydrolysis of ADP-ribose-arginine bond by GST-ADP-ribosylarginine hydrolase can be used to detect the arginine-ADP-ribosylated proteins in crude preparations. Arginine--ADP-ribosylated proteins in crude preparations. Arginine-ADP-ribosylated proteins in mouse spleen lymphocytes were identified using this method. << Less

  Anal Biochem 231:115-122(1995) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Molecular and immunological characterization of ADP-ribosylarginine hydrolases.

  Moss J., Stanley S.J., Nightingale M.S., Murtagh J.J. Jr., Monaco L., Mishima K., Chen H.C., Williamson K.C., Tsai S.C.

  Mono-ADP-ribosylation is a reversible modification of proteins with NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyzing the forward and reverse reactions, respectively. Hydrolase activities were present in a variety of animal species, with the highest specific activi ... >> More

  Mono-ADP-ribosylation is a reversible modification of proteins with NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyzing the forward and reverse reactions, respectively. Hydrolase activities were present in a variety of animal species, with the highest specific activities found in rat and mouse brain, spleen, and testis. Rat and mouse hydrolases were dithiothreitol- and Mg(2+)-dependent, whereas the bovine and guinea pig enzymes were dithiothreitol-independent. A rat brain hydrolase was purified approximately 20,000-fold and represented the major approximately 39-kDa protein on denaturing gels. Immunoaffinity-purified rabbit polyclonal antibodies reacted with 39-kDa proteins from turkey erythrocytes and rat, mouse, and calf brains. A rat brain cDNA library was screened using oligonucleotide and polymerase chain reaction-generated cDNA probes. Inserts from two overlapping clones yielded a composite sequence that included a 1086-base pair open reading frame, which contained amino acid sequences found in the purified hydrolase. A hydrolase fusion protein, synthesized in Escherichia coli, reacted with anti-39-kDa polyclonal antibodies and exhibited Mg(2+)- and dithiothreitol-dependent hydrolase activity. A coding region cDNA hybridized readily to a 1.7-kilobase band in rat and mouse poly(A)+ RNA, but poorly to bovine, chicken, rabbit, and human poly(A)+ RNA. The immunological and molecular biological data are consistent with partial conservation of hydrolase structure across animal species. << Less

  J. Biol. Chem. 267:10481-10488(1992) [PubMed] [EuropePMC]

• Identification of critical, conserved vicinal aspartate residues in mammalian and bacterial ADP-ribosylarginine hydrolases.

  Konczalik P., Moss J.

  NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyze opposing arms of a putative ADP-ribosylation cycle. ADP-ribosylarginine hydrolases from mammalian tissues and Rhodospirillum rubrum exhibit three regions of similarity in deduced amino acid sequence. We postulated tha ... >> More

  NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyze opposing arms of a putative ADP-ribosylation cycle. ADP-ribosylarginine hydrolases from mammalian tissues and Rhodospirillum rubrum exhibit three regions of similarity in deduced amino acid sequence. We postulated that amino acids in these consensus regions could be critical for hydrolase function. To test this hypothesis, hydrolase, cloned from rat brain, was expressed as a glutathione S-transferase fusion protein in Escherichia coli and purified by glutathione-Sepharose affinity chromatography. Conserved amino acids in each of these regions were altered by site-directed mutagenesis. Replacement of Asp-60 or Asp-61 with Ala, Gln, or Asn, but not Glu, significantly reduced enzyme activity. The double Asp-60 --> Glu/Asp-61 --> Glu mutant was inactive, as were Asp-60 --> Gln/Asp-61 --> Gln or Asp-60 --> Asn/Asp-61 --> Asn. The catalytically inactive single and double mutants appeared to retain conformation, since they bound ADP-ribose, a substrate analogue and an inhibitor of enzyme activity, with affinity similar to that of the wild-type hydrolase and with the expected stoichiometry of one. Replacing His-65, Arg-139, Asp-285, which are also located in the conserved regions, with alanine did not change specific activity. These data clearly show that the conserved vicinal aspartates 60 and 61 in rat ADP-ribosylarginine hydrolase are critical for catalytic activity, but not for high affinity binding of the substrate analogue, ADP-ribose. << Less

  J Biol Chem 274:16736-16740(1999) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Cloning and site-directed mutagenesis of human ADP-ribosylarginine hydrolase.

  Takada T., Iida K., Moss J.

  Mono-ADP-ribosylation of arginine is a reversible modification of proteins with NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyzing the opposing reactions in the cycle. ADP-ribosylarginine hydrolases differ in their dithiothreitol (DTT) requirements. Rat and mouse hy ... >> More

  Mono-ADP-ribosylation of arginine is a reversible modification of proteins with NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases catalyzing the opposing reactions in the cycle. ADP-ribosylarginine hydrolases differ in their dithiothreitol (DTT) requirements. Rat and mouse hydrolases require DTT for maximal activity, but calf, guinea pig, and human hydrolases are DTT-independent. To define the molecular basis for these differences, brain ADP-ribosylarginine hydrolases were cloned. Deduced amino acid sequences of mouse and rat hydrolases were 94% identical with 5 conserved cysteines. The human hydrolase sequence was 83% identical to that of rat but contained only 4 cysteines with cysteine 108 in rat corresponding to serine 103 in human. To investigate the role of rat cysteine 108, human and rat wild-type hydrolases and mutants, in which serine 103 in human was replaced by cysteine (S103C) and cysteine 108 in rat was replaced by serine (C108S), were expressed in Escherichia coli. Affinity-purified anti-rat brain hydrolase antibodies reacted with recombinant wild-type rat hydrolase, but only weakly with the C108S mutant. They did not react with human wild-type or the S103C mutant. Human hydrolase and rat C108S were DTT-independent; human S103C was, however, DTT-dependent. These data clearly show that cysteine 108 in rat hydrolase plays a critical role in DTT dependence and may be important in immunoreactivity. << Less

  J. Biol. Chem. 268:17837-17843(1993) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.