Structural and Functional Characterization of the Zn(II) Site in Dimethylargininase-1 (DDAH-1) from Bovine Brain

Knipp, Markus; Charnock, John; Garner, C. David; Vašák, Milan

doi:10.1074/jbc.m104056200

Cited by 46 publications

(62 citation statements)

References 60 publications

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“…Addition of zinc to DDAH preparations has consistently resulted in the inhibition of ADMA hydrolysis as reported in at least three studies (24,32,40). However, these studies used supraphysiological zinc concentrations and/or a purified or heavily diluted source of DDAH, so we sought to test more physiologically achievable zinc concentrations.…”

Section: Discussionmentioning

confidence: 80%

Contribution of whole blood to the control of plasma asymmetrical dimethylarginine

Billecke¹,

Kitzmiller²,

Northrup³

et al. 2006

American Journal of Physiology-Heart and Circulatory Physiology

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The endogenous nitric oxide (NO) synthase (NOS) inhibitor asymmetrical dimethylarginine (ADMA) is elevated in many patients and may contribute to the initiation and progression of their disease. While some mechanistic pathways have been identified, tissue-specific contributions to ADMA control remain unclear. We sought to determine if whole blood (WB) could participate in ADMA control ex vivo. Anesthetized male Sprague-Dawley rats underwent exsanguinations, and WB preparations were incubated at 37°C for 5 h. ADMA and symmetrical dimethylarginine were analyzed by high-pressure liquid chromatography. Incubation of lysed red blood cell (RBC) supernatant yielded a significant decrease in ADMA that was blocked by 4124W, a synthetic inhibitor of dimethylarginine dimethylaminohydrolase, the only reported enzyme to hydrolyze ADMA. Hydrolysis of ADMA was diminished by addition of physiologically relevant concentrations of zinc (i.e., 20 M). Conversely, when rat WB or WB supernatant was incubated at 37°C, it liberated quantities of free ADMA (1-2 M) that in vivo would likely have pathological consequences. Addition of arginine methyltransferase inhibitors to these incubations did not reduce ADMA release, indicating no dominant role for active protein methylation during these incubations. This ADMA liberation was significantly reduced by addition of protease inhibitors, indicating a dependence on peptide bond hydrolysis. Total ADMA (protein incorporated plus free) was determined by acid hydrolysis and found to be 43.18 Ϯ 4.79 M in WB with ϳ95% of this in RBCs.These ex vivo data demonstrate the potential of blood to control the NO-NOS system by modulating free ADMA. nitric oxide; protein arginine methyltransferase; symmetrical dimethylarginine; protease NITRIC OXIDE (NO) is produced by NO synthase (NOS) and has a myriad of functions throughout the body, including vasodilation, signaling functions, and inhibition of platelet aggregation. Deficiency or loss of NOS activity has been termed "endothelial dysfunction" and may contribute to a spectrum of cardiovascular diseases. Asymmetrical dimethylarginine (ADMA) and N G -monomethyl-L-arginine (L-NMMA) are endogenous inhibitors of NOS, but the concentration of ADMA exceeds that of L-NMMA by ϳ10-fold (60) and hence is of greater potential clinical interest. ADMA and its noninhibitory regioisomer, symmetrical dimethylarginine (SDMA) (6, 12), are released into the plasma following the breakdown of proteins containing arginine residues previously dimethylated by protein arginine methyltransferases (PRMT) (37). The literature contains compelling evidence that elevated plasma ADMA exists in diabetes mellitus (37), hypertension (1, 15, 22, 45, 51, 52), hypercholesterolemia (6), hyperhomocyst(e)inemia (50), experimental hemorrhage (4), preeclampsia (16), and sickle cell anemia (47). ADMA has also been reported to increase platelet activation and atherogenesis in the cardiovascular system (33). Cooke (11, 13) and others (17,59) give extensive reviews of much of these data and argue that ...

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Section: Discussionmentioning

confidence: 80%

Contribution of whole blood to the control of plasma asymmetrical dimethylarginine

Billecke¹,

Kitzmiller²,

Northrup³

et al. 2006

American Journal of Physiology-Heart and Circulatory Physiology

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“…(93,122,123,188). Knipp et al report that DDAH-1 is a Zn(II)-containing protein (23,80). Zn(II) is not involved in the catalytic process, but it is required to stabilize the enzyme in a fully active form (80).…”

Section: Brief Overview Of Development Of Knowledge On Ddahmentioning

confidence: 99%

“…In buffers that lack an appreciable metal-binding affinity, the effects of pH and temperature on enzyme activity reflect the presence of a small amount of Zn(II)-free DDAH-1. The Zn(II)-free enzyme has maximal activity at physiological pH (80).…”

Section: Brief Overview Of Development Of Knowledge On Ddahmentioning

confidence: 99%

Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems

Palm

Onozato²,

Luo³

et al. 2007

American Journal of Physiology-Heart and Circulatory Physiology

286

301

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Palm F, Onozato ML, Luo Z, Wilcox CS. Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems. Am J Physiol Heart Circ Physiol 293: H3227-H3245, 2007. First published October 12, 2007; doi:10.1152 doi:10. /ajpheart.00998.2007 )-dimethylarginine (ADMA) inhibits nitric oxide (NO) synthases (NOS). ADMA is a risk factor for endothelial dysfunction, cardiovascular mortality, and progression of chronic kidney disease. Two isoforms of dimethylarginine dimethylaminohydrolase (DDAH) metabolize ADMA. DDAH-1 is the predominant isoform in the proximal tubules of the kidney and in the liver. These organs extract ADMA from the circulation. DDAH-2 is the predominant isoform in the vasculature, where it is found in endothelial cells adjacent to the cell membrane and in intracellular vesicles and in vascular smooth muscle cells among the myofibrils and the nuclear envelope. In vivo gene silencing of DDAH-1 in the rat and DDAH ϩ/Ϫ mice both have increased circulating ADMA, whereas gene silencing of DDAH-2 reduces vascular NO generation and endothelium-derived relaxation factor responses. DDAH-2 also is expressed in the kidney in the macula densa and distal nephron. Angiotensin type 1 receptor activation in kidneys reduces the expression of DDAH-1 but increases the expression of DDAH-2. This rapidly evolving evidence of isoform-specific distribution and regulation of DDAH expression in the kidney and blood vessels provides potential mechanisms for nephron site-specific regulation of NO production. In this review, the recent advances in the regulation and function of DDAH enzymes, their roles in the regulation of NO generation, and their possible contribution to endothelial dysfunction in patients with cardiovascular and kidney diseases are discussed. nitric oxide synthase; hypertension; diabetes mellitus; chronic kidney disease; asymmetric dimethylarginineis an endogenous methylated amino acid that inhibits the constitutive endothelial (e) or type III and neuronal (n) or type I isoforms of nitric oxide (NO) synthase (NOS) (49,91,103,199). It is a less potent inhibitor of the inducible (i) or type II NOS isoform (41,191,213). Proteins are subject to methylation of arginine residues by protein arginine methyltransferase (PRMT). S-adenosylmethionine, which is synthesized from methionine and ATP, serves as the methyl donor and, in the process, is converted to S-adenosylhomocysteine, which itself can be hydrolyzed to homocysteine. Remethylation of homocysteine in the "remethylation pathway" regenerates methionine (14,179). ADMA is released by protein hydrolysis and exported from the cell and taken up by other cells via system y ϩ carriers of the cationic amino acid (CAT) family (14,196,212). ADMA is eliminated both by renal excretion and metabolic degradation. Its metabolism is facilitated by dimethylarginine dimethylaminohydrolases (DDAHs), which are expressed as type 1 and 2 isoforms. Recent studies have shown differential sites of expression of DDAH-1 and -2 in blo...

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“…23 A requirement for zinc has been identified for a bovine DDAH, but it is not clear whether this is a general requirement for DDAHs. 26,27 DDAHs are highly conserved throughout evolution 28 and have been identified in primitive organisms including bacteria. In higher organisms, including humans, 2 isoforms of DDAH have been identified that are encoded by genes located on chromosomes 1 (DDAH-1) and 6 (DDAH-2).…”

Section: Degredation Of Adma: the Ddahsmentioning

confidence: 99%

Cardiovascular Biology of the Asymmetric Dimethylarginine:Dimethylarginine Dimethylaminohydrolase Pathway

Vallance

Leiper

2004

ATVB

511

380

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Abstract-An increasing number of reports indicate that endogenously produced inhibitors of nitric oxide synthase, particularly asymmetric dimethylarginine (ADMA), regulate nitric oxide generation in disease states. This article describes the biology of ADMA and the implications for cardiovascular physiology and pathophysiology. Key Words: asymmetric dimethylarginine Ⅲ dimethylarginine dimethylaminohydrolase Ⅲ nitric oxide synthase Ⅲ endothelial dysfunction A symmetric dimethylarginine (ADMA) is a naturally occurring amino acid that circulates in plasma, is excreted in urine, and is found in tissues and cells. [1][2][3] It has aroused interest because it inhibits nitric oxide synthases (NOSs) 1 and therefore has the potential to produce considerable biological effects, particularly in the cardiovascular system. Recently, several studies have suggested that the plasma concentrations of ADMA provide a marker of risk for endothelial dysfunction and cardiovascular disease. 1,4 -6 This article describes the biology of ADMA and the implications for cardiovascular physiology and pathophysiology. How Is ADMA Made?ADMA is synthesized when arginine residues in proteins are methylated by the action of protein arginine methyltransferases (PRMTs). 7,8 Protein arginine methylation is a posttranslational modification that adds either 1 or 2 methyl groups to the guanidine nitrogens of arginine incorporated into proteins. There are 2 broad types of PRMTs: type 1 catalyze the formation of ADMA, whereas type 2 methylate both of the guanidino nitrogens and so result in the formation of symmetric dimethylarginine (SDMA; Figure 1). Both types of PRMT, of which there are several isoforms, can also monomethylate, leading to the formation of N G -monomethyl-L-arginine (L-NMMA). 7,8 Once the proteins are hydrolyzed, free methylarginines appear in the cytosol. The asymmetrically methylated arginines (ADMA and L-NMMA) are inhibitors of NOS, whereas SDMA is not. There is potentially a very broad range of substrate proteins for type 1 PRMTs, 9 and the enzymes and their substrates are widely distributed throughout the body. 10 The role of protein arginine methylation is unclear, but this process has been implicated in regulation of RNA binding, transcriptional regulation, DNA repair, protein localization, protein-protein interaction, signal transduction, and recycling or desensitization of receptors. However, it is only after the proteins are degraded that free methylarginines appear in the cytosol; to date, no direct route of synthesizing ADMA from free arginine has been identified. Thus, the amount of ADMA generated within a cell is dependent on the extent of arginine methylation in proteins and the rates of protein turnover.Because of the complex process leading to generation of free ADMA, it is unclear whether ADMA generation is fairly constant, it alters with PRMT activity, or if rates of protein turnover are the most important influence. Recently, studies with relatively nonspecific and low-potency PRMT inhibitors have suggested that PR...

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Structural and Functional Characterization of the Zn(II) Site in Dimethylargininase-1 (DDAH-1) from Bovine Brain

Cited by 46 publications

References 60 publications

Contribution of whole blood to the control of plasma asymmetrical dimethylarginine

Contribution of whole blood to the control of plasma asymmetrical dimethylarginine

Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems

Cardiovascular Biology of the Asymmetric Dimethylarginine:Dimethylarginine Dimethylaminohydrolase Pathway

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