Exogenous nitric oxide suppresses K<sub>ATP</sub> channel activity through S-nitrosylation_Journal of Biomedical Engineering

Authors：

WANG Xuefei ^1,2 , ZHANG Yuxin ¹ , HU Weicao ¹ ,  HAO Xuewei ^1,3

1. Harbin Medical University-Daqing, Daqing, Heilongjiang 163319, P. R. China;
2. The Fifth Affiliated Hospital of Herbin Medical University, Daqing, Heilongjiang 163319, P. R. China;
3. Engineering Technology Research Center for Precision Diagnosis and Treatment of Frigid Zone-Related Diseases in Heilongjiang Province, Daqing, Heilongjiang 163319, P. R. China;

Corresponding?author：

HAO Xuewei, Email: 382637958@qq.com

Keywords：

K_ATP; Nitric oxide; S-nitrosylation; S-glutathionylation

DOI：

10.7507/1001-5515.202511038

Video：

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The inhibitory effect of exogenous nitric oxide (NO) on the open state of ATP-sensitive potassium channels (K_ATP) and its underlying mechanism remain unclear. In this study, patch-clamp and molecular biology techniques are used to investigate this issue. In acutely isolated rat mesenteric artery smooth muscle cells and human embryonic kidney 293 cells (HEK293) expressing inwardly rectifying potassium channel 6.1 subunit/sulfonylurea receptor 2B subunit (Kir6.1/SUR2B), sodium nitroprusside (SNP) was found to significantly inhibit the activity of open K_ATP channels. Detection using biotin-labeled glutathione ethyl ester (BioGEE) combined with Western blotting showed that Kir6.1 subunit glutathionylation level was significantly decreased after SNP treatment. These results indicate that exogenous NO directly inhibits the activity of open K_ATP channels by nitrosylating key cysteine residues of the Kir6.1 subunit and competitively inhibiting glutathionylation at this site. This study provides new experimental evidence for the molecular mechanism of NO in vascular regulation.

Citation： WANG Xuefei, ZHANG Yuxin, HU Weicao, HAO Xuewei. Exogenous nitric oxide suppresses K_ATP channel activity through S-nitrosylation. Journal of Biomedical Engineering, 2026, 43(2): 391-397, 404. doi: 10.7507/1001-5515.202511038 Copy

1.	Hewat T I, Yau D, Jerome J C S, et al. Birth weight and diazoxide unresponsiveness strongly predict the likelihood of congenital hyperinsulinism due to a mutation in ABCC8 or KCNJ11. Eur J Endocrinol, 2021, 185(6): 813-818..
2.	Noma A. ATP-regulated K+ channels in cardiac muscle. Nature, 1983, 305(5930): 147-148..
3.	Clement A, Christensen S L, Jansen-Olesen I, et al. The ATP sensitive potassium channel (K_ATP) is a novel target for migraine drug development. Front Mol Neurosci, 2023, 16: 1182515..
4.	Martin G M, Patton B L, Shyng S L. K_ATP channels in focus: progress toward a structural understanding of ligand regulation. Curr Opin Struct Biol, 2023, 79: 102541..
5.	Nichols C G. Personalized therapeutics for K_ATP-dependent pathologies. Annu Rev Pharmacol Toxicol, 2023, 63: 541-563..
6.	Campbell J D, Sansom M S, Ashcroft F M. Potassium channel regulation. EMBO Rep, 2003, 4(11): 1038-1042..
7.	Lee S J, Maeda S, Gao J, et al. Oxidation driven reversal of PIP₂-dependent gating in GIRK₂ channels. Function (Oxf), 2023, 4(3): zqad016..
8.	Musaogullari A, Chai Y C. Redox regulation by protein S-glutathionylation: from molecular mechanisms to implications in health and disease. Int J Mol Sci, 2020, 21(21): 8113..
9.	Rashdan N A, Shrestha B, Pattillo C B. S-glutathionylation, friend or foe in cardiovascular health and disease. Redox Biol, 2020, 37: 101693..
10.	Rosas P C, Solaro R J. Implications of S-glutathionylation of sarcomere proteins in cardiac disorders, therapies, and diagnosis. Front Cardiovasc Med, 2022, 9: 1060716..
11.	Alcock L J, Perkins M V, Chalker J M. Chemical methods for mapping cysteine oxidation. Chem Soc Rev, 2018, 47(1): 231-268..
12.	Percio A, Cicchinelli M, Masci D, et al. Oxidative cysteine post translational modifications drive the redox code underlying neurodegeneration and amyotrophic lateral sclerosis. Antioxidants (Basel), 2024, 13(8): 883..
13.	Mao C, Zhao J, Cheng N, et al. S-nitrosylation in cardiovascular disorders: the state of the art. Biomolecules, 2025, 15(8): 1073..
14.	Feng J, Chen L, Zuo J. Protein S-nitrosylation in plants: current progresses and challenges. J Integr Plant Biol, 2019, 61(12): 1206-1223..
15.	Shi X, Qiu H. Post-translational S-nitrosylation of proteins in regulating cardiac oxidative stress. Antioxidants (Basel), 2020, 9(11): 1051..
16.	Murphy E, Kohr M, Menazza S, et al. Signaling by S-nitrosylation in the heart. J Mol Cell Cardiol, 2014, 73: 18-25..
17.	Monica F Z, Antunes E. Stimulators and activators of soluble guanylate cyclase for urogenital disorders. Nat Rev Urol, 2018, 15(1): 42-54..
18.	Krishnan S M, Kraehling J R, Eitner F, et al. The impact of the nitric oxide (NO)/soluble guanylyl cyclase (sGC) signaling cascade on kidney health and disease: a preclinical perspective. Int J Mol Sci, 2018, 19(6): 1712..
19.	Griffiths K, Lee J J, Frenneaux M P, et al. Nitrite and myocardial ischaemia reperfusion injury. where are we now? Pharmacol Ther, 2021, 223: 107819..
20.	張峰, 王志鵬, 王汝濤, 等. 一氧化氮誘導心肌細胞預適應早期保護作用的信號轉導途徑. 中國臨床藥理學與治療學, 2004(7): 766-769..
21.	Qu Z, Zhang J, Gao W, et al. Vasorelaxant effects of cerebralcare granule^? are mediated by NO/cGMP pathway, potassium channel opening and calcium channel blockade in isolated rat thoracic aorta. J Ethnopharmacol, 2014, 155(1): 572-579..
22.	Yang Y, Shi W, Cui N, et al. Oxidative stress inhibits vascular K_ATP channels by S-glutathionylation. J Biol Chem, 2010, 285(49): 38641-38648..
23.	Yu L, Jin X, Yang Y, et al. Rosiglitazone inhibits vascular K_ATP channels and coronary vasodilation produced by isoprenaline. Br J Pharmacol, 2011, 164(8): 2064-2072..
24.	Ashcroft F M. KATP channels and the metabolic regulation of insulin secretion in health and disease: the 2022 Banting Medal for scientific achievement award lecture. Diabetes, 2023, 72(6): 693-702..
25.	Rutter G A, Sweet I R. Glucose regulation of β-cell K_ATP channels: is a new model needed? Diabetes, 2024, 73(6): 849-855..
26.	Kim H J, Norton C E, Zawieja S D, et al. Acute metabolic stress induces lymphatic dysfunction through K_ATP channel activation. Function (Oxf), 2024, 5(5): zqae033..
27.	Driggers C M, Kuo Y Y, Zhu P, et al. Structure of an open K_ATP channel reveals tandem PIP₂ binding sites mediating the Kir6. 2 and SUR1 regulatory interface. Nat Commun, 2024, 15(1): 2502..
28.	Shi Y, Cui N, Shi W, et al. A short motif in Kir6. 1 consisting of four phosphorylation repeats underlies the vascular K_ATP channel inhibition by protein kinase C. J Biol Chem, 2008, 283(5): 2488-2494..
29.	Jiao J, Garg V, Yang B, et al. Protein kinase C-epsilon induces caveolin-dependent internalization of vascular adenosine 5'-triphosphate-sensitive K⁺ channels. Hypertension, 2008, 52(3): 499-506..
30.	Quinn K V, Giblin J P, Tinker A. Multisite phosphorylation mechanism for protein kinase A activation of the smooth muscle ATP-sensitive K⁺ channel. Circ Res, 2004, 94(10): 1359-1366..
31.	Shi Y, Wu Z, Cui N, et al. PKA phosphorylation of SUR2B subunit underscores vascular K_ATP channel activation by beta-adrenergic receptors. Am J Physiol Regul Integr Comp Physiol, 2007, 293(3): R1205-R1214..
32.	Gurrieri L, Capuzzi A C, Müller-Schüssele S J, et al. Dynamic regulation of arabidopsis β-amylase1 by glutathione and thioredoxins affects starch in guard cells. Plant Physiol, 2025, 198(4): kiaf344..
33.	Yang Y, Shi W, Chen X, et al. Molecular basis and structural insight of vascular KATP channel gating by S-glutathionylation. J Biol Chem, 2011, 286(11): 9298-9307..
34.	Kawano T, Zoga V, Kimura M, et al. Nitric oxide activates ATP-sensitive potassium channels in mammalian sensory neurons: action by direct S-nitrosylation. Mol Pain, 2009, 5: 12..
35.	崔琪, 杜雄, 校蕾, 等. 轉錄因子的巰基亞硝基化修飾及其生理學意義. 生理科學進展, 2023, 54(1): 26-32..

1. Hewat T I, Yau D, Jerome J C S, et al. Birth weight and diazoxide unresponsiveness strongly predict the likelihood of congenital hyperinsulinism due to a mutation in ABCC8 or KCNJ11. Eur J Endocrinol, 2021, 185(6): 813-818..
2. Noma A. ATP-regulated K+ channels in cardiac muscle. Nature, 1983, 305(5930): 147-148..
3. Clement A, Christensen S L, Jansen-Olesen I, et al. The ATP sensitive potassium channel (K_ATP) is a novel target for migraine drug development. Front Mol Neurosci, 2023, 16: 1182515..
4. Martin G M, Patton B L, Shyng S L. K_ATP channels in focus: progress toward a structural understanding of ligand regulation. Curr Opin Struct Biol, 2023, 79: 102541..
5. Nichols C G. Personalized therapeutics for K_ATP-dependent pathologies. Annu Rev Pharmacol Toxicol, 2023, 63: 541-563..
6. Campbell J D, Sansom M S, Ashcroft F M. Potassium channel regulation. EMBO Rep, 2003, 4(11): 1038-1042..
7. Lee S J, Maeda S, Gao J, et al. Oxidation driven reversal of PIP₂-dependent gating in GIRK₂ channels. Function (Oxf), 2023, 4(3): zqad016..
8. Musaogullari A, Chai Y C. Redox regulation by protein S-glutathionylation: from molecular mechanisms to implications in health and disease. Int J Mol Sci, 2020, 21(21): 8113..
9. Rashdan N A, Shrestha B, Pattillo C B. S-glutathionylation, friend or foe in cardiovascular health and disease. Redox Biol, 2020, 37: 101693..
10. Rosas P C, Solaro R J. Implications of S-glutathionylation of sarcomere proteins in cardiac disorders, therapies, and diagnosis. Front Cardiovasc Med, 2022, 9: 1060716..
11. Alcock L J, Perkins M V, Chalker J M. Chemical methods for mapping cysteine oxidation. Chem Soc Rev, 2018, 47(1): 231-268..
12. Percio A, Cicchinelli M, Masci D, et al. Oxidative cysteine post translational modifications drive the redox code underlying neurodegeneration and amyotrophic lateral sclerosis. Antioxidants (Basel), 2024, 13(8): 883..
13. Mao C, Zhao J, Cheng N, et al. S-nitrosylation in cardiovascular disorders: the state of the art. Biomolecules, 2025, 15(8): 1073..
14. Feng J, Chen L, Zuo J. Protein S-nitrosylation in plants: current progresses and challenges. J Integr Plant Biol, 2019, 61(12): 1206-1223..
15. Shi X, Qiu H. Post-translational S-nitrosylation of proteins in regulating cardiac oxidative stress. Antioxidants (Basel), 2020, 9(11): 1051..
16. Murphy E, Kohr M, Menazza S, et al. Signaling by S-nitrosylation in the heart. J Mol Cell Cardiol, 2014, 73: 18-25..
17. Monica F Z, Antunes E. Stimulators and activators of soluble guanylate cyclase for urogenital disorders. Nat Rev Urol, 2018, 15(1): 42-54..
18. Krishnan S M, Kraehling J R, Eitner F, et al. The impact of the nitric oxide (NO)/soluble guanylyl cyclase (sGC) signaling cascade on kidney health and disease: a preclinical perspective. Int J Mol Sci, 2018, 19(6): 1712..
19. Griffiths K, Lee J J, Frenneaux M P, et al. Nitrite and myocardial ischaemia reperfusion injury. where are we now? Pharmacol Ther, 2021, 223: 107819..
20. 張峰, 王志鵬, 王汝濤, 等. 一氧化氮誘導心肌細胞預適應早期保護作用的信號轉導途徑. 中國臨床藥理學與治療學, 2004(7): 766-769..
21. Qu Z, Zhang J, Gao W, et al. Vasorelaxant effects of cerebralcare granule^? are mediated by NO/cGMP pathway, potassium channel opening and calcium channel blockade in isolated rat thoracic aorta. J Ethnopharmacol, 2014, 155(1): 572-579..
22. Yang Y, Shi W, Cui N, et al. Oxidative stress inhibits vascular K_ATP channels by S-glutathionylation. J Biol Chem, 2010, 285(49): 38641-38648..
23. Yu L, Jin X, Yang Y, et al. Rosiglitazone inhibits vascular K_ATP channels and coronary vasodilation produced by isoprenaline. Br J Pharmacol, 2011, 164(8): 2064-2072..
24. Ashcroft F M. KATP channels and the metabolic regulation of insulin secretion in health and disease: the 2022 Banting Medal for scientific achievement award lecture. Diabetes, 2023, 72(6): 693-702..
25. Rutter G A, Sweet I R. Glucose regulation of β-cell K_ATP channels: is a new model needed? Diabetes, 2024, 73(6): 849-855..
26. Kim H J, Norton C E, Zawieja S D, et al. Acute metabolic stress induces lymphatic dysfunction through K_ATP channel activation. Function (Oxf), 2024, 5(5): zqae033..
27. Driggers C M, Kuo Y Y, Zhu P, et al. Structure of an open K_ATP channel reveals tandem PIP₂ binding sites mediating the Kir6. 2 and SUR1 regulatory interface. Nat Commun, 2024, 15(1): 2502..
28. Shi Y, Cui N, Shi W, et al. A short motif in Kir6. 1 consisting of four phosphorylation repeats underlies the vascular K_ATP channel inhibition by protein kinase C. J Biol Chem, 2008, 283(5): 2488-2494..
29. Jiao J, Garg V, Yang B, et al. Protein kinase C-epsilon induces caveolin-dependent internalization of vascular adenosine 5'-triphosphate-sensitive K⁺ channels. Hypertension, 2008, 52(3): 499-506..
30. Quinn K V, Giblin J P, Tinker A. Multisite phosphorylation mechanism for protein kinase A activation of the smooth muscle ATP-sensitive K⁺ channel. Circ Res, 2004, 94(10): 1359-1366..
31. Shi Y, Wu Z, Cui N, et al. PKA phosphorylation of SUR2B subunit underscores vascular K_ATP channel activation by beta-adrenergic receptors. Am J Physiol Regul Integr Comp Physiol, 2007, 293(3): R1205-R1214..
32. Gurrieri L, Capuzzi A C, Müller-Schüssele S J, et al. Dynamic regulation of arabidopsis β-amylase1 by glutathione and thioredoxins affects starch in guard cells. Plant Physiol, 2025, 198(4): kiaf344..
33. Yang Y, Shi W, Chen X, et al. Molecular basis and structural insight of vascular KATP channel gating by S-glutathionylation. J Biol Chem, 2011, 286(11): 9298-9307..
34. Kawano T, Zoga V, Kimura M, et al. Nitric oxide activates ATP-sensitive potassium channels in mammalian sensory neurons: action by direct S-nitrosylation. Mol Pain, 2009, 5: 12..
35. 崔琪, 杜雄, 校蕾, 等. 轉錄因子的巰基亞硝基化修飾及其生理學意義. 生理科學進展, 2023, 54(1): 26-32..

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Exogenous nitric oxide suppresses K_ATP channel activity through S-nitrosylation

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Journal of Biomedical Engineering

Exogenous nitric oxide suppresses KATP channel activity through S-nitrosylation

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Exogenous nitric oxide suppresses K_ATP channel activity through S-nitrosylation