Characterization of antibody against human liver guanase by immunoblotting and immunohistochemical staining of human liver guanase by a direct labeling antibody-enzyme method.

Ito, Susumo; Iwasaki, Akiharu; Syundo, Jyoji; Tamura, Yoshiyuki; Kishi, Seiichiro; Mori, Hiromu; Ii, Kunio; Matsuda, Yoshiko; Katsunuma, N

doi:10.1177/37.5.2649557

Cited by 6 publications

(5 citation statements)

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“…GAH catalyzes the hydrolytic deamination of guanine, yielding xanthine and ammonium, and is considered to be involved in a major pathway for producing uric acid (30). As consistent with our findings, GAH was shown to be abundant in brain, and its activity was detected in both nuclear and cytosolic fractions (31)(32)(33), although the biological significance has been largely unknown.…”

Section: Discussionsupporting

confidence: 78%

A Novel NE-dlg/SAP102-associated Protein, p51-nedasin, Related to the Amidohydrolase Superfamily, Interferes with the Association between NE-dlg/SAP102 and N-Methyl-d-aspartate Receptor

Kuwahara¹,

Araki²,

Makino³

et al. 1999

Journal of Biological Chemistry

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The membrane-associated guanylate kinase proteins have been known to interact various membrane receptors with their N-terminal segments designated the PDZ domains and to cluster these receptors at the target site of the cell membrane. NE-dlg/SAP102, a neuronal and endocrine tissue-specific MAGUK family protein, was found to be expressed in both dendrites and cell bodies in neuronal cells. Although NE-dlg/SAP102 localized at dendrites was shown to interact with N-methyl-D-aspartate receptor 2B via the PDZ domains to compose postsynaptic density, the binding proteins existing in the cell body of the neuron are still unknown. Here we report the isolation of a novel NE-dlg/SAP102-associated protein, p51-nedasin. Nedasin has a significant homology with amidohydrolase superfamily proteins and shows identical sequences to a recently identified protein that has guanine aminohydrolase activity. Nedasin has four alternative splice variants (S, V1, V2, and V3) that exhibited different C-terminal structures. NE-dlg/ SAP102 is shown to interact with only the S form of nedasin which is predominantly expressed in brain. The expression of nedasin in neuronal cells increases in parallel with the progress of synaptogenesis and is mainly detected in cell bodies where it co-localizes with NE-dlg/ SAP102. Furthermore, nedasin interferes with the association between NE-dlg/SAP102 and NMDA receptor 2B in vitro. These findings suggest that alternative splicing of nedasin may play a role in the formation and/or structural change in synapses during neuronal development by modifying clustering of neurotransmitter receptors at the synaptic sites.At the sites of cell-cell contacts of epithelial cells or the synaptic junctions of neuronal cells, several membrane receptors and channels are clustered into multiprotein complexes linked to the cytoskeleton via interactions of their C-terminal cytoplasmic tails with a novel protein family called membraneassociated guanylate kinase homologues (MAGUK) 1 (1). The MAGUK family proteins contain three distinct domains as follows: an N-terminal segment comprised of one or three copies of an 80 -90-amino acid motif called the PDZ (PSD-95/Dlg/ ZO-1) domain, an src homology 3 (SH3) domain, and a region with high similarity to guanylate kinases (GK) (2, 3). The PDZ domain is utilized as a module for interacting with the Cterminal Xaa-(Ser/Thr)-Xaa-Val (X(S/T)XV) motif of various proteins and generating multiprotein complexes (4, 5).Each MAGUK protein is thought to perform a distinct function depending upon its tissue distribution, cellular localization, and associated molecules. For instance, PSD-95/SAP90, which is one of the MAGUK proteins, is predominantly expressed in the brain and localizes at the postsynaptic membrane and presynaptic axon terminals of inhibitory neurons (6 -8). PSD-95/SAP90 binds to the cytoplasmic tail of both Shaker-type voltage-gated K ϩ channels and the 2B subunit of N-methyl-D-aspartate (NMDA)-type glutamate receptors (7, 9 -11). PSD-95/SAP90 expressed in neuronal cells is therefo...

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Section: Discussionsupporting

confidence: 78%

A Novel NE-dlg/SAP102-associated Protein, p51-nedasin, Related to the Amidohydrolase Superfamily, Interferes with the Association between NE-dlg/SAP102 and N-Methyl-d-aspartate Receptor

Kuwahara¹,

Araki²,

Makino³

et al. 1999

Journal of Biological Chemistry

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“…In a previous study (13), the immunohistochemical reaction was positive at the same sites as the histochemical reaction for guanase with DAB in human liver sections and was negative in control sections. Thus the distribution of the brown discoloration of oxidized DAB obtained by this method was in accordance with the specific location of guanase.…”

Section: Discussionmentioning

confidence: 99%

“…We used the purified enzyme to prepare antihuman liver guanase serum. Then we studied the distribution of guanase in human kidney by a previously reported histochemical staining method (7) and by the direct labelling antibody enzyme method (13). We also examined the guanase activity in nephron segments by a micromethod (19).…”

Section: Discussionmentioning

confidence: 99%

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Histochemical and biochemical studies on guanase in the human and rat kidney.

Ito

Maeda

Iwasaki

et al. 1991

Acta Histochem. Cytochem

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A specific antibody against purified human liver guanase was raised in rabbits. The antiserum formed a single precipitin line with human liver extracts, and completely inhibited the activity of the enzyme derived from human liver and kidney. Immunoblotting showed that the antibody bound only to the one protein band of liver and kidney guanase activity. In immunohistochemical studies, the DAB (3, 3'-diaminobenzidine tetrahydrochloride)-reaction with this antibody was positive in the same locations as the histochemical guanase reaction. Moreover, biochemical data of enzymic activity in nephron segments of the rat indicated that guanase activity was located in the proximal tubules. Thus, the DAB reaction appeared to show the location of guanase itself. This immunohistochemical procedure for detection of guanase should useful in clinical studies on this enzyme.

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“…In contrast, histochemical and immunohistochemical analyses of human kidney and liver assigned cytoplasmic localisation [Ito et al, 1989;Kubo et al, 2006], which was also shown by differential centrifugation in rat and mouse brain cells [Kumar et al, 1972].…”

Section: ) Was Homologous To the Conserved Sequence Pgx[v/i] Dxh[t/mentioning

confidence: 98%

<b><i>Arxula adeninivorans</i></b> Recombinant Guanine Deaminase and Its Application in the Production of Food with Low Purine Content

Trautwein-Schult

Jankowska²,

Cordes³

et al. 2014

Microb Physiol

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Purines of exogenous and endogenous sources are degraded to uric acid in human beings. Concentrations >6.8 mg uric acid/dl serum cause hyperuricemia and its symptoms. Pharmaceuticals and the reduction of the intake of purine-rich food are used to control uric acid levels. A novel approach to the latter proposition is the enzymatic reduction of the purine content of food by purine-degrading enzymes. Here we describe the production of recombinant guanine deaminase by the yeast Arxula adeninivorans LS3 and its application in food. In media supplemented with nitrogen sources hypoxanthine or adenine, guanine deaminase (AGDA) gene expression is induced and intracellular accumulation of guanine deaminase (Agdap) protein occurs. The characteristics of the guanine deaminase isolated from wild-type strain LS3 and a transgenic strain expressing the AGDA gene under control of the strong constitutive TEF1 promoter were determined and compared. Both enzymes were dimeric and had temperature optima of 55°C with high substrate specificity for guanine and localisation in both the cytoplasm and vacuole of yeast. The enzyme was demonstrated to reduce levels of guanine in food. A mixture of guanine deaminase and other purine degradation enzymes will allow the reduction of purines in purine-rich foods.

show abstract

Characterization of antibody against human liver guanase by immunoblotting and immunohistochemical staining of human liver guanase by a direct labeling antibody-enzyme method.

Cited by 6 publications

References 4 publications

A Novel NE-dlg/SAP102-associated Protein, p51-nedasin, Related to the Amidohydrolase Superfamily, Interferes with the Association between NE-dlg/SAP102 and N-Methyl-d-aspartate Receptor

A Novel NE-dlg/SAP102-associated Protein, p51-nedasin, Related to the Amidohydrolase Superfamily, Interferes with the Association between NE-dlg/SAP102 and N-Methyl-d-aspartate Receptor

Histochemical and biochemical studies on guanase in the human and rat kidney.

<b><i>Arxula adeninivorans</i></b> Recombinant Guanine Deaminase and Its Application in the Production of Food with Low Purine Content

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