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Single atom substitution in mouse protein kinase G eliminates oxidant sensing to cause hypertension

Abstract

Blood pressure regulation is crucial for the maintenance of health, and hypertension is a risk factor for myocardial infarction, heart failure, stroke and renal disease. Nitric oxide (NO) and prostacyclin trigger well-defined vasodilator pathways; however, substantial vasorelaxation in response to agents such as acetylcholine persists when the synthesis of these molecules is prevented. This remaining vasorelaxation activity, termed endothelium-derived hyperpolarizing factor (EDHF), is more prevalent in resistance than in conduit blood vessels and is considered a major mechanism for blood pressure control1,2,3,4. Hydrogen peroxide (H2O2) has been shown to be a major component of EDHF in several vascular beds in multiple species, including in humans5,6,7,8,9,10. H2O2 causes the formation of a disulfide bond between the two α subunits of protein kinase G I-α (PKGI-α), which activates the kinase independently of the NO–cyclic guanosine monophosphate (cGMP) pathway and is coupled to vasodilation11. To test the importance of PKGI-α oxidation in the EDHF mechanism and blood pressure control in vivo, we generated a knock-in mouse expressing only a C42S 'redox-dead' version of PKGI-α. This amino acid substitution, a single-atom change (an oxygen atom replacing a sulfur atom), blocked the vasodilatory action of H2O2 on resistance vessels and resulted in hypertension in vivo.

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Figure 1: Differential responses of WT and knock-in vessels to oxidant interventions.
Figure 2: Differential responses of WT and knock-in vessels to acetylcholine.
Figure 3: Knock-in mice are hypertensive compared to WT littermates.
Figure 4: Comparison of cardiac output, vascular fibrosis and renal function in WT and knock-in mice.

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Acknowledgements

We acknowledge support from the Medical Research Council, the British Heart Foundation, the Leducq Foundation and the UK Department of Health through the National Institute for Health Research comprehensive Biomedical Research Centre award to Guy's & St Thomas' National Health Service Foundation Trust.

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Authors

Contributions

O.P. conducted the myography and protein analysis experiments and wrote the manuscript. O.R. undertook in vivo blood pressure and inulin clearance studies and wrote the manuscript. P.E. supervised the project and wrote the manuscript.

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Correspondence to Philip Eaton.

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The authors declare no competing financial interests.

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Supplementary Figures 1–7 and Supplementary Table 1 (PDF 448 kb)

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Prysyazhna, O., Rudyk, O. & Eaton, P. Single atom substitution in mouse protein kinase G eliminates oxidant sensing to cause hypertension. Nat Med 18, 286–290 (2012). https://doi.org/10.1038/nm.2603

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