Publications

• Guanidino acid hydrolysis by the human enzyme annotated as agmatinase.

  Sinn M., Stanoppi M., Hauth F., Fleming J.R., Funck D., Mayans O., Hartig J.S.

  Guanidino acids such as taurocyamine, guanidinobutyrate, guanidinopropionate, and guanidinoacetate have been detected in humans. However, except for guanidionacetate, which is a precursor of creatine, their metabolism and potential functions remain poorly understood. Agmatine has received consider ... >> More

  Guanidino acids such as taurocyamine, guanidinobutyrate, guanidinopropionate, and guanidinoacetate have been detected in humans. However, except for guanidionacetate, which is a precursor of creatine, their metabolism and potential functions remain poorly understood. Agmatine has received considerable attention as a potential neurotransmitter and the human enzyme so far annotated as agmatinase (AGMAT) has been proposed as an important modulator of agmatine levels. However, conclusive evidence for the assigned enzymatic activity is lacking. Here we show that AGMAT hydrolyzed a range of linear guanidino acids but was virtually inactive with agmatine. Structural modelling and direct biochemical assays indicated that two naturally occurring variants differ in their substrate preferences. A negatively charged group in the substrate at the end opposing the guanidine moiety was essential for efficient catalysis, explaining why agmatine was not hydrolyzed. We suggest to rename AGMAT as guanidino acid hydrolase (GDAH). Additionally, we demonstrate that the GDAH substrates taurocyamine, guanidinobutyrate and guanidinopropionate were produced by human glycine amidinotransferase (GATM). The presented findings show for the first time an enzymatic activity for GDAH/AGMAT. Since agmatine has frequently been proposed as an endogenous neurotransmitter, the current findings clarify important aspects of the metabolism of agmatine and guanidino acid derivatives in humans. << Less

  Sci. Rep. 12:22088-22088(2022) [PubMed] [EuropePMC]

  This publication is cited by 6 other entries.

• The purification and characterization of human kidney L-arginine:glycine amidinotransferase.

  Gross M.D., Eggen M.A., Simon A.M., Van Pilsum J.F.

  Human kidney L-arginine:glycine amidinotransferase (transamidinase) has been purified to a homogeneous state as defined by native and sodium dodecyl sulfate gel electrophoresis and by ultracentrifugation (sedimentation equilibrium) experiments. The four steps in the isolation procedure were chroma ... >> More

  Human kidney L-arginine:glycine amidinotransferase (transamidinase) has been purified to a homogeneous state as defined by native and sodium dodecyl sulfate gel electrophoresis and by ultracentrifugation (sedimentation equilibrium) experiments. The four steps in the isolation procedure were chromatography with DEAE-cellulose, gel filtration with Sephadex G-150, chromatography with phenyl Sepharose, and high-pressure liquid chromatography with hydroxylapatite. The final product represented a 90-fold purification of the enzyme. Human kidney transamidinase is a dimer with a molecular mass of 89,000 Da and subunit masses of 44,000 Da. The Km for arginine and glycine were both 2.5 mM and the Vmax was 0.5 mumol ornithine/min/mg protein. The ultraviolet absorption spectrum, specific activity, and isoelectric points were determined for human kidney transamidinase. Multiple forms of the enzyme were obtained by isoelectric focusing. Human kidney transamidinase cross-reacted with polyclonal antibodies raised to rat kidney transamidinase. All of the properties of human kidney transamidinase that we have examined were similar to those of rat kidney transamidinase. A close evolutionary relationship between the rat and human kidney transamidinase is suggested. << Less

  Arch. Biochem. Biophys. 251:747-755(1986) [PubMed] [EuropePMC]

• A novel superfamily of enzymes that catalyze the modification of guanidino groups.

  Shirai H., Blundell T.L., Mizuguchi K.

  Three enzymes, peptidyl-arginine deiminase from Porphyromonas gingivalis, arginine deiminase and amidinotransferase are traditionally classified separately. By combining PSI-BLAST and FUGUE, data presented in this article describe how these enzymes belong to a novel superfamily, adopting a common ... >> More

  Three enzymes, peptidyl-arginine deiminase from Porphyromonas gingivalis, arginine deiminase and amidinotransferase are traditionally classified separately. By combining PSI-BLAST and FUGUE, data presented in this article describe how these enzymes belong to a novel superfamily, adopting a common fold and sharing similar catalytic mechanisms. << Less

  Trends Biochem Sci 26:465-468(2001) [PubMed] [EuropePMC]

• Recombinant expression and isolation of human L-arginine:glycine amidinotransferase and identification of its active-site cysteine residue.

  Humm A., Fritsche E., Mann K., Goehl U., Huber R.

  Creatine and its phosphorylated form play a central role in the energy metabolism of muscle and nerve tissues. l-Arginine:glycine amidinotransferase (AT) catalyses the committed step in the formation of creatine. The mitochondrial and cytosolic forms of the enzyme are believed to derive from the s ... >> More

  Creatine and its phosphorylated form play a central role in the energy metabolism of muscle and nerve tissues. l-Arginine:glycine amidinotransferase (AT) catalyses the committed step in the formation of creatine. The mitochondrial and cytosolic forms of the enzyme are believed to derive from the same gene by alternative splicing. We have expressed recombinant human AT in Escherichia coli with two different N-termini, resembling the longest two forms of the enzyme that we had isolated recently from porcine kidney mitochondria as a mixture. The enzymes were expressed with N-terminal histidine tags followed by factor Xa-cleavage sites. We established a new method for the removal of N-terminal fusion peptides by means of an immobilized snake venom prothrombin activator. We identified cysteine-407 as the active-site residue of AT by radioactive labelling and isolation of labelled peptides, and by site-directed mutagenesis of the protein. << Less

  Biochem. J. 322:771-776(1997) [PubMed] [EuropePMC]

• The effect of growth hormone and thyroxine on the amount of L-arginine:glycine amidinotransferase in kidneys of hypophysectomized rats. Purification and some properties of rat kidney transamidinase.

  McGuire D.M., Tormanen C.D., Segal I.S., Van Pilsum J.F.

  J. Biol. Chem. 255:1152-1159(1980) [PubMed] [EuropePMC]

• Synthesis of neuroactive guanidino compounds by rat kidney L-arginine: glycine amidinotransferase.

  Watanabe Y., Van Pilsum J.F., Yokoi I., Mori A.

  Several neuroactive guanidino compounds have been reported to be synthesized in mammals by transamidination reactions. The enzyme(s) responsible for their synthesis and their location in the body has not been well established. The purpose of this investigation was to determine if purified homogene ... >> More

  Several neuroactive guanidino compounds have been reported to be synthesized in mammals by transamidination reactions. The enzyme(s) responsible for their synthesis and their location in the body has not been well established. The purpose of this investigation was to determine if purified homogeneous rat kidney alpha- and beta-L-arginine : glycine amidinotransferase (transamidinase) would catalyze the synthesis of certain neuroactive guanidino compounds, and if so, to determine if any catalytic specificity existed between the two forms of the enzymes. L-Arginine (Arg) was used as the amidino group donor and the following compounds were investigated for their ability to accept the amidino group: ethanolamine; 4-aminobutyric acid; lysine; 5-aminovaleric acid; 3-aminopropionic acid; taurine; L-glutamic acid (Glu); L-aspartic acid (Asp); and histidine (His). All of the above listed compounds served as amidino group acceptors for the enzyme except Glu, Asp and His. No differences were found between the alpha- and beta-transamidinase in any of the experiments reported, and the synthesis of 2-guanidinoethanol by the enzyme was by a sequential mechanism with a Km for Arg and ethanolamine of 14mM and 163mM, respectively. The possibility that the site of synthesis of the neuroactive guanidino compounds in the kidney and perhaps pancreas is discussed. << Less

  Life Sci. 55:351-358(1994) [PubMed] [EuropePMC]

• Crystal structure and mechanism of human L-arginine:glycine amidinotransferase: a mitochondrial enzyme involved in creatine biosynthesis.

  Humm A., Fritsche E., Steinbacher S., Huber R.

  L-arginine:glycine amidinotransferase (AT) catalyses the committed step in creatine biosynthesis by formation of guanidinoacetic acid, the immediate precursor of creatine. We have determined the crystal structure of the recombinant human enzyme by multiple isomorphous replacement at 1.9 A resoluti ... >> More

  L-arginine:glycine amidinotransferase (AT) catalyses the committed step in creatine biosynthesis by formation of guanidinoacetic acid, the immediate precursor of creatine. We have determined the crystal structure of the recombinant human enzyme by multiple isomorphous replacement at 1.9 A resolution. A telluromethionine derivative was used in sequence assignment. The structure of AT reveals a new fold with 5-fold pseudosymmetry of circularly arranged betabeta alphabeta-modules. These enclose the active site compartment, which is accessible only through a narrow channel. The overall structure resembles a basket with handles that are formed from insertions into the betabeta alphabeta-modules. Binding of L-ornithine, a product inhibitor, reveals a marked induced-fit mechanism, with a loop at the active site entrance changing its conformation accompanied by a shift of an alpha-helix by -4 A. Binding of the arginine educt to the inactive mutant C407A shows a similar mode of binding. A reaction mechanism with a catalytic triad Cys-His-Asp is proposed on the basis of substrate and product bound states. << Less

  EMBO J. 16:3373-3385(1997) [PubMed] [EuropePMC]