Potassium Effects on NCC Are Attenuated during Inhibition of Cullin E3-Ubiquitin Ligases.
Potassium Effects on NCC Are Attenuated during Inhibition of Cullin E3-Ubiquitin Ligases.
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DOI：
10.3390/cells11010095
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发表时间：
2021-12-29
期刊：
影响因子：
6
通讯作者：
Fenton RA
中科院分区：
文献类型：
--
作者：
Murali SK;Little R;Poulsen SB;Ferdaus MZ;Ellison DH;McCormick JA;Fenton RA
噻嗪敏感性氯化钠协同转运蛋白（NCC）在维持钠（Na+）和钾（K+）稳态中起着至关重要的作用。NCC活性由无赖氨酸激酶1和4（WNK 1和WNK 4）调节，其丰度由RING型E3连接酶Cullin 3（Cul 3）及其底物接头Kelch样蛋白3控制。膳食K+摄入量与NCC活性呈负相关，但这一现象背后的机制仍有待充分阐明。在这里，我们研究了参与介导K+对NCC磷酸化（活性形式）和丰度的影响的cullin家族的其他成员。在不同的K+含量的饮食喂养的小鼠肾脏中，NCC（磷酸化和总）和活性（neddylated）形式的cullin（Cul 1，3，4和5）之间存在负相关性。在Cul 3突变小鼠（CUL 3-Het/Δ9）中，高饮食K+对磷酸化NCC的影响减弱。短期（30分钟）和长期（24小时）的细胞外K+浓度的变化不影响cullin neddylation水平在离体肾小管。短期内，在存在MLN 4924（泛cullin抑制剂）的情况下，高细胞外K+降低NCC磷酸化的能力得以保留，但对低细胞外K+无反应。从长期来看，MLN 4924减弱了高细胞外K+对NCC磷酸化的影响，并且不存在对低细胞外K+的反应。我们的数据表明，除了Cul 3，其他cullin参与介导K+对NCC磷酸化和丰度的影响。
The thiazide-sensitive sodium chloride cotransporter (NCC) plays a vital role in maintaining sodium (Na+) and potassium (K+) homeostasis. NCC activity is modulated by with-no-lysine kinases 1 and 4 (WNK1 and WNK4), the abundance of which is controlled by the RING-type E3 ligase Cullin 3 (Cul3) and its substrate adapter Kelch-like protein 3. Dietary K+ intake has an inverse correlation with NCC activity, but the mechanism underlying this phenomenon remains to be fully elucidated. Here, we investigated the involvement of other members of the cullin family in mediating K+ effects on NCC phosphorylation (active form) and abundance. In kidneys from mice fed diets varying in K+ content, there were negative correlations between NCC (phosphorylated and total) and active (neddylated) forms of cullins (Cul1, 3, 4, and 5). High dietary K+ effects on phosphorylated NCC were attenuated in Cul3 mutant mice (CUL3-Het/Δ9). Short-term (30 min) and long-term (24 h) alterations in the extracellular K+ concentration did not affect cullin neddylation levels in ex vivo renal tubules. In the short term, the ability of high extracellular K+ to decrease NCC phosphorylation was preserved in the presence of MLN4924 (pan-cullin inhibitor), but the response to low extracellular K+ was absent. In the long term, MLN4924 attenuated the effects of high extracellular K+ on NCC phosphorylation, and responses to low extracellular K+ were absent. Our data suggest that in addition to Cul3, other cullins are involved in mediating the effects of K+ on NCC phosphorylation and abundance.