Increased Nitration of Sarcoplasmic Reticulum Ca
            <sup>2+</sup>
            -ATPase in Human Heart Failure

Lokuta, Andrew J.; Maertz, Nathan A.; Meethal, Sivan Vadakkadath; Potter, Katherine T.; Kamp, Timothy J.; Valdivia, Héctor H.; Haworth, Robert A.

doi:10.1161/01.cir.0000156461.81529.d7

Cited by 131 publications

(104 citation statements)

References 62 publications

(60 reference statements)

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“…The capacity of ILK to co-IP with SERCA-2a and PLN in transgenic myocardium ( Supplementary Figs 1 and 3) and the direct correlation between the protein but not mRNA expression levels of ILK and SERCA-2a following modulated ILK expression in iPSC-CMs ( Supplementary Fig. 2 ARTICLE post-translational modifications including nitration 55 . Overexpression of ILK, using for example gene therapy, would nevertheless be expected to increase SERCA-2a/PLN function and thereby improve cardiac function in DCM through a proteinbinding-dependent mechanism, irrespective of the primary inciting process.…”

Section: Discussionmentioning

confidence: 96%

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Traister

Aafaqi

et al. 2014

Nat Commun

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Human dilated cardiomyopathy (DCM) manifests as a profound reduction in biventricular cardiac function that typically progresses to death or cardiac transplantation. There is no effective mechanism-based therapy currently available for DCM, in part because the transduction of mechanical load into dynamic changes in cardiac contractility (termed mechanotransduction) remains an incompletely understood process during both normal cardiac function and in disease states. Here we show that the mechanoreceptor protein integrin-linked kinase (ILK) mediates cardiomyocyte force transduction through regulation of the key calcium regulatory protein sarcoplasmic/endoplasmic reticulum Ca 2 þ ATPase isoform 2a (SERCA-2a) and phosphorylation of phospholamban (PLN) in the human heart. A non-oncogenic ILK mutation with a synthetic point mutation in the pleckstrin homology-like domain (ILK R211A ) is shown to enhance global cardiac function through SERCA-2a/PLN. Thus, ILK serves to link mechanoreception to the dynamic modulation of cardiac contractility through a previously undiscovered interaction with the functional SERCA-2a/PLN module that can be exploited to rescue impaired mechanotransduction in DCM.

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Section: Discussionmentioning

confidence: 96%

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Traister

Aafaqi

et al. 2014

Nat Commun

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“…Increased tyrosine nitration is another indicator of oxidative/nitrosative damage to SERCA2a protein (21,27,44). Therefore, we assessed the effects of Hsp70 on the formation of nitrotyrosine in SERCA2a molecules in response to heat stress.…”

Section: Expression Of Serca2a and Hsp70 In Hek-293 Cellsmentioning

confidence: 99%

Protective effects of Hsp70 on the structure and function of SERCA2a expressed in HEK-293 cells during heat stress

Fu¹,

Tupling²

2009

American Journal of Physiology-Heart and Circulatory Physiology

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Fu MH, Tupling AR. Protective effects of Hsp70 on the structure and function of SERCA2a expressed in HEK-293 cells during heat stress. Am J Physiol Heart Circ Physiol 296: H1175-H1183, 2009. First published February 27, 2009 doi:10.1152/ajpheart.01276.2008.-Heat shock protein 70 (Hsp70) can physically interact with and prevent thermal inactivation of sarco(endo)plasmic reticulum Ca 2ϩ -ATPase (SERCA) 1a, the SERCA isoform expressed in adult fasttwitch skeletal muscle. This study examined whether Hsp70 could physically interact with and prevent thermal inactivation of SERCA2a, the SERCA isoform expressed in heart. HEK-293 cells were cotransfected with cDNAs encoding human Hsp70 and rabbit SERCA2a (S2a/Hsp70). Cells cotransfected with SERCA2a cDNA and pMT2 (S2a/pMT2) were used as control. One-half of the cells was heat shocked at 40°C for 1 h (HS), and one-half was maintained at 37°C before harvesting the cells and isolating microsomes. Western blot analysis showed that Hsp70 and SERCA2a were colocalized in the microsomal fraction. The levels of Hsp70 were approximately fivefold higher (P Ͻ 0.05) in S2a/Hsp70 compared with S2a/pMT2 and approximately twofold higher (P Ͻ 0.05) following HS in all cells. Coimmunoprecipitation demonstrated that Hsp70 directly binds to SERCA2a. Following HS, maximal SERCA2a activity was reduced (ϳ52%, P Ͻ 0.05) in S2a/pMT2 but was increased (ϳ33%, P Ͻ 0.05) in S2a/Hsp70. Thermal inactivation of SERCA2a in S2a/pMT2 was associated with decreased (ϳ49%, P Ͻ 0.05) binding capacity for fluorescein isothiocyanate (FITC) and increased carbonyl (ϳ42%, P Ͻ 0.05) and nitrotyrosine (ϳ40%, P Ͻ 0.05) levels in SERCA2a. By contrast, the HS-induced increase in maximal SERCA2a activity observed in S2a/Hsp70 corresponded with no change (P Ͼ 0.05) in FITC-binding capacity and reductions in carbonyl (ϳ40%, P Ͻ 0.05) and nitrotyrosine (ϳ23%, P Ͻ 0.05) levels in SERCA2a compared with S2a/pMT2. These results show that Hsp70 forms a protective interaction with SERCA2a during HS actually reducing oxidation and nitrosylation of SERCA2a thus increasing its maximal activity. coimmunoprecipitatoin; carbonyl; nitrotyrosine; nucleotide binding site SARCO(ENDO)PLASMIC reticulum Ca 2ϩ -ATPases (SERCAs) are typical of the class of P-type ATPases that bind specific ions to be transported, hydrolyze ATP to form a phosphoprotein intermediate, and undergo conformational changes resulting in ion translocation (29). SERCA pumps are 110-kDa integral membrane proteins that consist of 10 transmembrane helexes (M1-M10), three cytoplasmic domains (A, actuator; N, nucleotide-binding; P, phosphorylation), and small luminal loops (48). In humans, 3 genes (ATP2A1-3) generate multiple isoforms (SERCAla,b, SERCA2a-c, SECA3a-f) by developmental or tissue-specific alternative splicing (14). Two SERCA isoforms predominate in adult striated muscle, namely SERCA1a and SERCA2a. The SERCA1a isoform consists of 994 amino acids and accounts for Ͼ99% of the SERCA isoforms expressed in adult fast-twitch skeletal muscle, whereas SERCA2a consists...

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“…Additionally, the lack of effect of low SIN-1 in PLB −/− myocytes may indicate that these positive effects are mediated via PLB-dependent mechanisms. However, the effects of low SIN-1 on other excitation-contraction coupling proteins cannot be completely ruled out, as peroxynitrite has been shown to have effects on other excitation-contraction coupling proteins [35][36][37][38][39]. Further, SIN-1 has a complex chemistry that could potentially lead to the formation of other reactive nitrogen species.…”

Section: Sin-1 and Plb −/− Myocyte Functionmentioning

confidence: 99%

Biphasic effect of SIN-1 is reliant upon cardiomyocyte contractile state

Kohr¹,

Wang²,

Wheeler³

et al. 2008

Free Radical Biology and Medicine

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Many studies have demonstrated a biphasic effect of peroxynitrite in the myocardium, but few studies have investigated this biphasic effect on β-adrenergic responsiveness and its dependence on contractile state. We have previously shown that high SIN-1 (source of peroxynitrite, 200 μmol/L) produced significant anti-adrenergic effects during maximal β-adrenergic stimulation in cardiomyocytes. In the current study, we hypothesize that the negative effects of high SIN-1 will be greatest during high contractile states, while the positive effects of low SIN-1 (10 μmol/L) will predominate during low contractility. Isolated murine cardiomyocytes were field stimulated at 1 Hz and [Ca 2+ ] i transients and shortening were recorded. Following submaximal ISO (β-adrenergic agonist, 0.01 μmol/L) stimulation, 200 μmol/L SIN-1 induced two distinct phenomena. Cardiomyocytes undergoing a large response to ISO showed a significant reduction in contractility, while cardiomyocytes exhibiting a modest response to ISO showed a further increase in contractility. Additionally, 10 μmol/L SIN-1 always increased contractility during low ISO stimulation, but had no effect during maximal ISO (1 μmol/L) stimulation. 10 μmol/L SIN-1 also increased basal contractility. Interestingly, SIN-1 only produced a contractile effect under one condition in phospholamban knockout cardiomyocytes, providing a potential mechanism for the biphasic effect of peroxynitrite. These results provide clear evidence for a biphasic effect of peroxynitrite, with high peroxynitrite modulating high levels of β-adrenergic responsiveness and low peroxynitrite regulating basal function and low levels of β-adrenergic stimulation.

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Increased Nitration of Sarcoplasmic Reticulum Ca ²⁺ -ATPase in Human Heart Failure

Cited by 131 publications

References 62 publications

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Protective effects of Hsp70 on the structure and function of SERCA2a expressed in HEK-293 cells during heat stress

Biphasic effect of SIN-1 is reliant upon cardiomyocyte contractile state

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Increased Nitration of Sarcoplasmic Reticulum Ca 2+ -ATPase in Human Heart Failure

Cited by 131 publications

References 62 publications

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function

Protective effects of Hsp70 on the structure and function of SERCA2a expressed in HEK-293 cells during heat stress

Biphasic effect of SIN-1 is reliant upon cardiomyocyte contractile state

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Increased Nitration of Sarcoplasmic Reticulum Ca ²⁺ -ATPase in Human Heart Failure