Purification and Characterization of Peptidylarginine Deiminase from Rabbit Skeletal Muscle

Takahara, Hidenari; Oikawa, Yoshihiro; Sugawara, Kiyoshi

doi:10.1093/oxfordjournals.jbchem.a134548

Cited by 57 publications

(38 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, we did not observe any synthesis of NO when the synaptosomal preparation was incubated with 200 kLM NH4Cl. Furthermore, Na-benzoyl-L-arginine ethyl ester, a substrate for peptidyl-arginine deiminase (20), was not a substrate, and formamidine, an inhibitor of bacterial arginine deiminase (21), had no effect in the present study. On the basis of its NADPH dependence, we have suggested that the reaction in vascular endothelial cells is an oxygenation (9).…”

Section: Discussioncontrasting

confidence: 63%

Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase.

Knowles

Palacios

Palmer

et al. 1989

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

1,210

610

View full text Add to dashboard Cite

A soluble enzyme obtained from rat forebrain catalyzes the NADPH-dependent formation of nitric oxide (NO) and citrulline from L-arginine. The NO formed stimulates the soluble guanylate cyclase and this stimulation is abolished by low concentrations of hemoglobin. The synthesis of NO and citrulline is dependent on the presence of physiological concentrations of free Ca2+ and is inhibited by NG-monomethyl-L-arginine, but not by its enantiomer NGmonomethylDarginin e or by Lcanavanine. L-Homoarginine, L-argyl-L-apartate, or L-arginine methyl ester can replace L-ginine as substrates for the enzyme. These results indicate that NO is formed from Larginine in the brain through an enzymic reaction similar to that in vascular endothelial cells, neutrophils, and macrophages, adding support to our hypothesis that the formation of NO from L-arginine is a widespread transduction mechanism for the stimulation of the soluble guanylate cyclase. Activated macrophages also synthesize NOj and NO3 from the terminal guanido nitrogen atom(s) of L-arginine (5). This reaction, which occurs via the formation of NO (6), is involved in the cytotoxic activities of these cells (7). We have demonstrated recently that rat peritoneal neutrophils form NO from L-arginine (8).The endothelial cell and macrophage enzyme that forms NO from L-arginine is soluble, is NADPH-dependent, forms citrulline as a coproduct, and is inhibited by NI-monomethyl-L-arginine refs. 6 and 9). Furthermore, in both cells the enzyme requires a divalent cation, which in the case of the macrophage has been suggested to be Mg2+ (6).Some years ago, L-arginine was identified as an endogenous activator of the soluble guanylate cyclase in brain tissue (10). Since this activation resembled that of the nitrovasodilators (11) and NO is known to stimulate soluble guanylate cyclase in the brain (12), we have investigated the existence in the central nervous system of an enzymic system capable of converting L-arginine into NO and citrulline.While this work was in progress Garthwaite et al. (13) Preparation of Crude Synaptosomal Cytosol. Male rats (200-300 g, four for each preparation) were killed by cervical dislocation and the forebrains were rapidly removed and cooled in ice-cold washing buffer (0.32 M sucrose/10 mM Hepes/0.1 mM EDTA, pH 7.4); subsequent procedures were carried out at 0-40C. The tissue was placed in fresh washing buffer, finely minced, washed once with 50 ml of washing buffer and twice with homogenization buffer (0.32 M sucrose/10 mM Hepes, 1 mM DL-dithiothreitol, pH 7.4) to remove contaminating erythrocytes, and homogenized with 20 strokes of a Dounce homogenizer. The homogenate was diluted to 50 ml with homogenization buffer and centrifuged (1400 x g, 10 min); the supernatant obtained was centrifuged (18,000 x g, 10 min) to obtain a crude synaptosomal pellet. After aspiration of the supernatant, 8 ml of 1 mM DLdithiothreitol (dissolved in distilled water) was added to the pellet to cause hypotonic swelling of the synaptosomes, which were then lysed by ho...

show abstract

Section: Discussioncontrasting

confidence: 63%

Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase.

Knowles

Palacios

Palmer

et al. 1989

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

1,210

610

View full text Add to dashboard Cite

show abstract

“…8 ) These properties were confirmed by this study showing the absolute requirements for calcium ion and dithiothreitol for the modification of contrapsin (Fig. 3).…”

Section: Discussionsupporting

confidence: 78%

Specific Modification of an Arginine Residue in Mouse Contrapsin by Peptidylarginine Deiminase Altered Its Inhibitory Activities against Trypsin and Chymotrypsin

Takahara

Sugawara

1987

Agricultural and Biological Chemistry

Self Cite

View full text Add to dashboard Cite

“…To determine whether PAD2 was increased in the white matter tracts of heterozygous PD2 mice, we used a polyclonal antibody generated against whole muscle PAD2 (Takahara et al, 1983;Takahara et al, 1989) to stain mouse brain sections. The antibody labeled white matter tracts in the corpus callosum of both normal and PD2 mice, with a higher degree of labeling in the white matter tracts of the PD2 brain ( Fig.…”

Section: Expression Of Pad2 Protein In Pd2 Micementioning

confidence: 99%

Peptidylarginine deiminase 2 (PAD2) overexpression in transgenic mice leads to myelin loss in the central nervous system

Musse¹,

Li²,

Ackerley³

et al. 2008

Disease Models &Amp; Mechanisms

123

View full text Add to dashboard Cite

SUMMARYDemyelination in the central nervous system is the hallmark feature in multiple sclerosis (MS). The mechanism resulting in destabilization of myelin is a complex multi-faceted process, part of which involves deimination of myelin basic protein (MBP). Deimination, the conversion of protein-bound arginine to citrulline, is mediated by the peptidylarginine deiminase (PAD) family of enzymes, of which the PAD2 and PAD4 isoforms are present in myelin. To test the hypothesis that PAD contributes to destabilization of myelin in MS, we developed a transgenic mouse line (PD2) containing multiple copies of the cDNA encoding PAD2, under the control of the MBP promoter. Using previously established criteria, clinical signs were more severe in PD2 mice than in their normal littermates. The increase in PAD2 expression and activity in white matter was demonstrated by immunohistochemistry, reverse transcriptase-PCR, enzyme activity assays, and increased deimination of MBP. Light and electron microscopy revealed more severe focal demyelination and thinner myelin in the PD2 homozygous mice compared with heterozygous PD2 mice. Quantitation of the disease-associated molecules GFAP and CD68, as measured by immunoslot blots, were indicative of astrocytosis and macrophage activation. Concurrently, elevated levels of the pro-inflammatory cytokine TNF-α and nuclear histone deimination support initiation of demyelination by increased PAD activity. These data support the hypothesis that elevated PAD levels in white matter represents an early change that precedes demyelination.

show abstract

Purification and Characterization of Peptidylarginine Deiminase from Rabbit Skeletal Muscle

Cited by 57 publications

References 0 publications

Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase.

Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase.

Specific Modification of an Arginine Residue in Mouse Contrapsin by Peptidylarginine Deiminase Altered Its Inhibitory Activities against Trypsin and Chymotrypsin

Peptidylarginine deiminase 2 (PAD2) overexpression in transgenic mice leads to myelin loss in the central nervous system

Contact Info

Product

Resources

About