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Research paper
Published online: 2020-02-25
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Comparison of the protein acetylome of endothelial cells upon shear flow and resveratrol treatment

Wun-Rong Lin, Chung-Jung Liu, Yaw-Syan Fu, Fu-An Li, Bin Huang
DOI: 10.5603/CJ.a2020.0029
·
Pubmed: 32104903

open access

Ahead of print
Original articles
Published online: 2020-02-25

Abstract

Background: Posttranslational acetylation/deacetylation known as the acetylome is important in regulating protein activity. Shear flow (SF) and resveratrol (RSV) are two stimuli that represent physical and chemical signal separately. The acetylome co-regulated by these two stimuli remain unclear. Methods: Human umbilical cord vein endothelial cells (HUVECs) were subjected to either SF of 12 dynes/cm2 or 10 μM RSV. The purified acetylated peptides were labeled by isobaric tags for relative and absolute quantitation (iTRAQ) analysis. The signaling cascades of the identified acetylome were predicted by ingenuity pathway analysis (IPA). Co-immunoprecipitation was applied to confirm the acetylation status of proteins. Results: Five groups of proteins showed an increased acetylation upon SF and RSV treatment. After algorithm, 628 proteins with increased acetylation and 22 proteins with decreased acetylation were identified in the SF acetylome. For the acetylome regulated by RSV, 145 proteins with increased acetylation and 23 proteins with decreased acetylation were identified. Compared these two acetylomes, 129 proteins with increased acetylation and 2 proteins with decreased acetylation were co-regulated by both SF and RSV treatments. IPA analysis showed that this co-regulated acetylome was involved in heat shock response, and the signals of eNOS, STAT3, JAK/STAT and ERK/MAPK. Co-immunoprecipitation analysis further confirmed the acetylated status of mitochondrial HSP60 and mitochondrial citrate synthase. Conclusions: This study indicated that physical signal is more complicated than chemical signal in the case of acetylome. The co-regulated proteins are worthy for further study in discussing synergetic effect between physical and chemical signal in cardioprotection.

Abstract

Background: Posttranslational acetylation/deacetylation known as the acetylome is important in regulating protein activity. Shear flow (SF) and resveratrol (RSV) are two stimuli that represent physical and chemical signal separately. The acetylome co-regulated by these two stimuli remain unclear. Methods: Human umbilical cord vein endothelial cells (HUVECs) were subjected to either SF of 12 dynes/cm2 or 10 μM RSV. The purified acetylated peptides were labeled by isobaric tags for relative and absolute quantitation (iTRAQ) analysis. The signaling cascades of the identified acetylome were predicted by ingenuity pathway analysis (IPA). Co-immunoprecipitation was applied to confirm the acetylation status of proteins. Results: Five groups of proteins showed an increased acetylation upon SF and RSV treatment. After algorithm, 628 proteins with increased acetylation and 22 proteins with decreased acetylation were identified in the SF acetylome. For the acetylome regulated by RSV, 145 proteins with increased acetylation and 23 proteins with decreased acetylation were identified. Compared these two acetylomes, 129 proteins with increased acetylation and 2 proteins with decreased acetylation were co-regulated by both SF and RSV treatments. IPA analysis showed that this co-regulated acetylome was involved in heat shock response, and the signals of eNOS, STAT3, JAK/STAT and ERK/MAPK. Co-immunoprecipitation analysis further confirmed the acetylated status of mitochondrial HSP60 and mitochondrial citrate synthase. Conclusions: This study indicated that physical signal is more complicated than chemical signal in the case of acetylome. The co-regulated proteins are worthy for further study in discussing synergetic effect between physical and chemical signal in cardioprotection.

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Keywords

acetylome, shear flow, resveratrol, iTRAQ, HSP60, ATP-citrate synthase

About this article
Title

Comparison of the protein acetylome of endothelial cells upon shear flow and resveratrol treatment

Journal

Cardiology Journal

Issue

Ahead of print

Article type

Research paper

Published online

2020-02-25

DOI

10.5603/CJ.a2020.0029

Pubmed

32104903

Keywords

acetylome
shear flow
resveratrol
iTRAQ
HSP60
ATP-citrate synthase

Authors

Wun-Rong Lin
Chung-Jung Liu
Yaw-Syan Fu
Fu-An Li
Bin Huang

References (25)
  1. Narita T, Weinert BT, Choudhary C, et al. Functions and mechanisms of non-histone protein acetylation. Nat Rev Mol Cell Biol. 2019; 20(3): 156–174.
  2. Huang Bo, Yang XD, Lamb A, et al. Posttranslational modifications of NF-kappaB: another layer of regulation for NF-kappaB signaling pathway. Cell Signal. 2010; 22(9): 1282–1290.
  3. D'Onofrio N, Servillo L, Balestrieri ML. SIRT1 and SIRT6 Signaling Pathways in Cardiovascular Disease Protection. Antioxid Redox Signal. 2018; 28(8): 711–732.
  4. Zullo A, Simone E, Grimaldi M, et al. Sirtuins as mediator of the anti-ageing effects of calorie restriction in skeletal and cardiac muscle. Int J Mol Sci. 2018; 19(4).
  5. Zecchin A, Pattarini L, Gutierrez MI, et al. Reversible acetylation regulates vascular endothelial growth factor receptor-2 activity. J Mol Cell Biol. 2014; 6(2): 116–127.
  6. Hsieh HJ, Liu CA, Huang B, et al. Shear-induced endothelial mechanotransduction: the interplay between reactive oxygen species (ROS) and nitric oxide (NO) and the pathophysiological implications. J Biomed Sci. 2014; 21: 3.
  7. Huang B, Chen CT, Chen CS, et al. Laminar shear flow increases hydrogen sulfide and activates a nitric oxide producing signaling cascade in endothelial cells. Biochem Biophys Res Commun. 2015; 464(4): 1254–1259.
  8. Liu J, Bi X, Chen T, et al. Shear stress regulates endothelial cell autophagy via redox regulation and Sirt1 expression. Cell Death Dis. 2015; 6: e1827.
  9. Wu LH, Chang HC, Ting PC, et al. Laminar shear stress promotes mitochondrial homeostasis in endothelial cells. J Cell Physiol. 2018; 233(6): 5058–5069.
  10. Go YM, Son DJu, Park H, et al. Disturbed flow enhances inflammatory signaling and atherogenesis by increasing thioredoxin-1 level in endothelial cell nuclei. PLoS One. 2014; 9(9): e108346.
  11. Wu JM, Wang ZR, Hsieh TC, et al. Mechanism of cardioprotection by resveratrol, a phenolic antioxidant present in red wine (Review). Int J Mol Med. 2001; 8: 3–17.
  12. Xia N, Förstermann U, Li H. Resveratrol and endothelial nitric oxide. Molecules. 2014; 19(10): 16102–16121.
  13. Chen T, Li J, Liu J, et al. Activation of SIRT3 by resveratrol ameliorates cardiac fibrosis and improves cardiac function via the TGF-β/Smad3 pathway. Am J Physiol Heart Circ Physiol. 2015; 308(5): H424–H434.
  14. Lin MC, et al. Hsing, CH, Li, FA, Rosuvastatin modulates the post-translational acetylome in endothelial cells. Acta Cardiol Sin. 2014; 30: 67–73.
  15. Wiese S, Reidegeld KA, Meyer HE, et al. Protein labeling by iTRAQ: a new tool for quantitative mass spectrometry in proteome research. Proteomics. 2007; 7(3): 340–350.
  16. Chen Z, Peng IC, Cui X, et al. Shear stress, SIRT1, and vascular homeostasis. Proc Natl Acad Sci U S A. 2010; 107(22): 10268–10273.
  17. Klinge CM, Blankenship KA, Risinger KE, et al. Resveratrol and estradiol rapidly activate MAPK signaling through estrogen receptors alpha and beta in endothelial cells. J Biol Chem. 2005; 280(9): 7460–7468.
  18. Cappello F, Zummo G. HSP60 expression during carcinogenesis: a molecular "proteus" of carcinogenesis? Cell Stress Chaperones. 2005; 10(4): 263–264.
  19. Xu Q, Schett G, Seitz CS, et al. Surface staining and cytotoxic activity of heat-shock protein 60 antibody in stressed aortic endothelial cells. Circ Res. 1994; 75(6): 1078–1085.
  20. Hochleitner BW, Hochleitner EO, Obrist P, et al. Fluid shear stress induces heat shock protein 60 expression in endothelial cells in vitro and in vivo. Arterioscler Thromb Vasc Biol. 2000; 20(3): 617–623.
  21. Shi L, Tu BP. Acetyl-CoA and the regulation of metabolism: mechanisms and consequences. Curr Opin Cell Biol. 2015; 33: 125–131.
  22. Infantino V, Iacobazzi V, Palmieri F, et al. ATP-citrate lyase is essential for macrophage inflammatory response. Biochem Biophys Res Commun. 2013; 440(1): 105–111.
  23. Doddaballapur A, Michalik KM, Manavski Y, et al. Laminar shear stress inhibits endothelial cell metabolism via KLF2-mediated repression of PFKFB3. Arterioscler Thromb Vasc Biol. 2015; 35(1): 137–145.
  24. Zhang Y, Cui G, Wang Y, et al. SIRT1 activation alleviates brain microvascular endothelial dysfunction in peroxisomal disorders. Int J Mol Med. 2019; 44(3): 995–1005.
  25. Ballermann BJ, Dardik A, Eng E, et al. Shear stress and the endothelium. Kidney Int Suppl. 1998; 67: S100–S108.

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