Medicine and Health Sciences Commons^™

Protein Carbonylation Of An Amino Acid Residue Of The Na/K‐Atpase Α1 Subunit Determines Na/K‐Atpase Signaling And Sodium Transport In Renal Proximal Tubular Cells, Yanling Yan, Anna P. Shapiro, Brahma R. Mopidevi, Muhammad Chaudhry, Kyle Maxwell, Steven T. Haller, Christopher A. Drummond, David J. Keendey, Jiang Tian, Deepak Malhorta, Zijian Xie, Joseph I. Shapiro Md, Jiang Liu

Muhammad Chaudhry

Background We have demonstrated that cardiotonic steroids, such as ouabain, signaling through the Na/K‐ATPase, regulate sodium reabsorption in the renal proximal tubule. By direct carbonylation modification of the Pro222 residue in the actuator (A) domain of pig Na/K‐ATPase α1 subunit, reactive oxygen species are required for ouabain‐stimulated Na/K‐ATPase/c‐Src signaling and subsequent regulation of active transepithelial ²²Na⁺ transport. In the present study we sought to determine the functional role of Pro222 carbonylation in Na/K‐ATPase signaling and sodium handling.

Methods and Results Stable pig α1 knockdown LLC‐PK1‐originated PY‐17 cells were rescued by expressing wild‐type rat α1 and rat α1 with …

Protein Carbonylation Of An Amino Acid Residue Of The Na/K‐Atpase Α1 Subunit Determines Na/K‐Atpase Signaling And Sodium Transport In Renal Proximal Tubular Cells, Yanling Yan, Anna P. Shapiro, Brahma R. Mopidevi, Muhammad Chaudhry, Kyle Maxwell, Steven T. Haller, Christopher A. Drummond, David J. Keendey, Jiang Tian, Deepak Malhorta, Zijian Xie, Joseph I. Shapiro Md, Jiang Liu

Jiang Liu

Background We have demonstrated that cardiotonic steroids, such as ouabain, signaling through the Na/K‐ATPase, regulate sodium reabsorption in the renal proximal tubule. By direct carbonylation modification of the Pro222 residue in the actuator (A) domain of pig Na/K‐ATPase α1 subunit, reactive oxygen species are required for ouabain‐stimulated Na/K‐ATPase/c‐Src signaling and subsequent regulation of active transepithelial ²²Na⁺ transport. In the present study we sought to determine the functional role of Pro222 carbonylation in Na/K‐ATPase signaling and sodium handling.

Methods and Results Stable pig α1 knockdown LLC‐PK1‐originated PY‐17 cells were rescued by expressing wild‐type rat α1 and rat α1 with …

Protein Carbonylation Of An Amino Acid Residue Of The Na/K‐Atpase Α1 Subunit Determines Na/K‐Atpase Signaling And Sodium Transport In Renal Proximal Tubular Cells, Yanling Yan, Anna P. Shapiro, Brahma R. Mopidevi, Muhammad Chaudhry, Kyle Maxwell, Steven T. Haller, Christopher A. Drummond, David J. Keendey, Jiang Tian, Deepak Malhorta, Zijian Xie, Joseph I. Shapiro Md, Jiang Liu

Zijian Xie

Background We have demonstrated that cardiotonic steroids, such as ouabain, signaling through the Na/K‐ATPase, regulate sodium reabsorption in the renal proximal tubule. By direct carbonylation modification of the Pro222 residue in the actuator (A) domain of pig Na/K‐ATPase α1 subunit, reactive oxygen species are required for ouabain‐stimulated Na/K‐ATPase/c‐Src signaling and subsequent regulation of active transepithelial ²²Na⁺ transport. In the present study we sought to determine the functional role of Pro222 carbonylation in Na/K‐ATPase signaling and sodium handling.

Methods and Results Stable pig α1 knockdown LLC‐PK1‐originated PY‐17 cells were rescued by expressing wild‐type rat α1 and rat α1 with …

Protein Carbonylation Of An Amino Acid Residue Of The Na/K‐Atpase Α1 Subunit Determines Na/K‐Atpase Signaling And Sodium Transport In Renal Proximal Tubular Cells, Yanling Yan, Anna P. Shapiro, Brahma R. Mopidevi, Muhammad Chaudhry, Kyle Maxwell, Steven T. Haller, Christopher A. Drummond, David J. Keendey, Jiang Tian, Deepak Malhorta, Zijian Xie, Joseph I. Shapiro Md, Jiang Liu

Yanling Yan

Background We have demonstrated that cardiotonic steroids, such as ouabain, signaling through the Na/K‐ATPase, regulate sodium reabsorption in the renal proximal tubule. By direct carbonylation modification of the Pro222 residue in the actuator (A) domain of pig Na/K‐ATPase α1 subunit, reactive oxygen species are required for ouabain‐stimulated Na/K‐ATPase/c‐Src signaling and subsequent regulation of active transepithelial ²²Na⁺ transport. In the present study we sought to determine the functional role of Pro222 carbonylation in Na/K‐ATPase signaling and sodium handling.

Methods and Results Stable pig α1 knockdown LLC‐PK1‐originated PY‐17 cells were rescued by expressing wild‐type rat α1 and rat α1 with …

The Physiological And Clinical Importance Of Sodium Potassium Atpase In Cardiovascular Diseases, Yanling Yan, Joseph I. Shapiro Md

Yanling Yan

The Na/K-ATPase has been extensively studied, but it is only recently that its role as a scaffolding and signaling protein has been identified. It has been identified that cardiotonic steroids (CTS) such as digitalis mediate signal transduction through the Na/K-ATPase in a process found to result in the generation of reactive oxygen species (ROS). As these ROS also appear capable of initiating this signal cascade, a feed forward amplification process has been postulated and subsequently implicated in some disease pathways including uremic cardiomyopathy.

Attenuation Of Na/K-Atpase Mediated Oxidant Amplification With Pnaktide Ameliorates Experimental Uremic Cardiomyopathy, Jiang Liu, Jiang Tian, Muhammad Chaudhry, Nader G. Abraham, Joseph I Shapiro

Nader G. Abraham

We have previously reported that the sodium potassium adenosine triphosphatase (Na/K-ATPase) can effect the amplification of reactive oxygen species. In this study, we examined whether attenuation of oxidant stress by antagonism of Na/K-ATPase oxidant amplification might ameliorate experimental uremic cardiomyopathy induced by partial nephrectomy (PNx). PNx induced the development of cardiac morphological and biochemical changes consistent with human uremic cardiomyopathy. Both inhibition of Na/K-ATPase oxidant amplification with pNaKtide and induction of heme oxygenase-1 (HO-1) with cobalt protoporphyrin (CoPP) markedly attenuated the development of phenotypical features of uremic cardiomyopathy. In a reversal study, administration of pNaKtide after the induction of uremic …

Attenuation Of Na/K-Atpase Mediated Oxidant Amplification With Pnaktide Ameliorates Experimental Uremic Cardiomyopathy, Jiang Liu, Jiang Tian, Muhammad Chaudhry, Nader G. Abraham, Joseph I Shapiro

Jiang Liu

We have previously reported that the sodium potassium adenosine triphosphatase (Na/K-ATPase) can effect the amplification of reactive oxygen species. In this study, we examined whether attenuation of oxidant stress by antagonism of Na/K-ATPase oxidant amplification might ameliorate experimental uremic cardiomyopathy induced by partial nephrectomy (PNx). PNx induced the development of cardiac morphological and biochemical changes consistent with human uremic cardiomyopathy. Both inhibition of Na/K-ATPase oxidant amplification with pNaKtide and induction of heme oxygenase-1 (HO-1) with cobalt protoporphyrin (CoPP) markedly attenuated the development of phenotypical features of uremic cardiomyopathy. In a reversal study, administration of pNaKtide after the induction of uremic …

Central Role For The Cardiotonic Steroid Marinobufagenin In The Pathogenesis Of Experimental Uremic Cardiomyopathy, David Kennedy, Sandeep Vetteth, Sankaridrug Periyasamy, Mohamed Kanj, Larisa Fedorova, Samer Khouri, M. Kahaleh, Zijian Xie, Deepak Malhotra, Nikolai Kolodkin, Edward Lakatta, Olga Fedorova, Alexei Bagrov, Joseph Shapiro

Zijian Xie

Patients with chronic renal failure develop a “uremic” cardiomyopathy characterized by diastolic dysfunction, cardiac hypertrophy, and systemic oxidant stress. Patients with chronic renal failure are also known to have increases in the circulating concentrations of the cardiotonic steroid marinobufagenin (MBG). On this background, we hypothesized that elevations in circulating MBG may be involved in the cardiomyopathy. First, we observed that administration of MBG (10 g/kg per day) for 4 weeks caused comparable increases in plasma MBG as partial nephrectomy at 4 weeks. MBG infusion caused increases in conscious blood pressure, cardiac weight, and the time constant for left ventricular relaxation …

Marinobufagenin Stimulates Fibroblast Collagen Production And Causes Fibrosis In Experimental Uremic Cardiomyopathy, Jihad Elkareh, David Kennedy, Belvadi Yashaswi, Sandeep Vetteth, Amjad Shidyak, Eric Kim, Sleiman Smaili, Sankaridrug Periyasamy, Imad Hariri, Larisa Fedorova, Jiang Liu, Liang Wu, M. Kahaleh, Zijian Xie, Deepak Malhotra, Olga Fedorova, Vladimir Kashkin, Alexei Bagrov, Joseph Shapiro

Jiang Liu

We have observed recently that experimental renal failure in the rat is accompanied by increases in circulating concentrations of the cardiotonic steroid, marinobufagenin (MBG), and substantial cardiac fibrosis. We performed the following studies to examine whether MBG might directly stimulate cardiac fibroblast collagen production. In vivo studies were performed using the 5/6th nephrectomy model of experimental renal failure (PNx), MBG infusion (MBG), PNx after immunization against MBG, and concomitant PNx and adrenalectomy. Physiological measurements with a Millar catheter and immunohistochemistry were performed. In vitro studies were then pursued with cultured isolated cardiac fibroblasts. We observed that PNx and MBG increased …

Marinobufagenin Stimulates Fibroblast Collagen Production And Causes Fibrosis In Experimental Uremic Cardiomyopathy, Jihad Elkareh, David Kennedy, Belvadi Yashaswi, Sandeep Vetteth, Amjad Shidyak, Eric Kim, Sleiman Smaili, Sankaridrug Periyasamy, Imad Hariri, Larisa Fedorova, Jiang Liu, Liang Wu, M. Kahaleh, Zijian Xie, Deepak Malhotra, Olga Fedorova, Vladimir Kashkin, Alexei Bagrov, Joseph Shapiro

Zijian Xie

We have observed recently that experimental renal failure in the rat is accompanied by increases in circulating concentrations of the cardiotonic steroid, marinobufagenin (MBG), and substantial cardiac fibrosis. We performed the following studies to examine whether MBG might directly stimulate cardiac fibroblast collagen production. In vivo studies were performed using the 5/6th nephrectomy model of experimental renal failure (PNx), MBG infusion (MBG), PNx after immunization against MBG, and concomitant PNx and adrenalectomy. Physiological measurements with a Millar catheter and immunohistochemistry were performed. In vitro studies were then pursued with cultured isolated cardiac fibroblasts. We observed that PNx and MBG increased …

Ouabain Decreases Sarco(Endo)Plasmic Reticulum Calcium Atpase Activity In Rat Hearts By A Process Involving Protein Oxidation, David Kennedy, Sandeep Vetteth, Miaorong Xie, Sankaridrug Periyasamy, Jingwei Xie, Chi Han, Venkatesha Basrur, Krishna Mutgi, Vladimir Fedorov, Deepak Malhotra, Joseph Shapiro

Jingwei Xie

The effect of cardiac glycosides to increase cardiac inotropy by altering Ca2+ cycling is well known but still poorly understood. The studies described in this report focus on defining the effects of ouabain signaling on sarcoplasmic reticulum Ca2+-ATPase function. Rat cardiac myocytes treated with 50 μM ouabain demonstrated substantial increases in systolic and diastolic Ca2+ concentrations. The recovery time constant for the Ca2+ transient, τ, was significantly prolonged by ouabain. Exposure to 10 μM H2O2, which causes an increase in intracellular reactive oxygen species similar to that of 50 μM ouabain, caused a similar increase in τ. Concurrent exposure to …