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首页> 外文期刊>The protein journal >Post-translational Regulation of Hexokinase Function and Protein Stability in the Aestivating Frog Xenopus laevis
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Post-translational Regulation of Hexokinase Function and Protein Stability in the Aestivating Frog Xenopus laevis

机译：在培养青蛙非洲爪蟾中己糖激酶功能和蛋白质稳定性的翻译后调控。

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Xenopus laevis endure substantial dehydration which can impose hypoxic stress due to impaired blood flow. Tissues may increase reliance on anaerobic glycolysis for energy production making the regulation of hexokinase (HK) important. We investigated the enzymatic properties and phosphorylation state of purified HK from the muscle of control and dehydrated (30 % total body water lost) frogs. Bioinformatic tools were also applied to analyze the structural implication of HK phosphorylation in silico. HK from the muscle of dehydrated frogs showed a significantly higher V-max (3.4-fold) and K-m for glucose (2.4-fold) compared with control HK but the Km for ATP was unaltered. HK from dehydrated frogs also showed greater phosphoserine content (20 % increase) and lower phosphothreonine (22 % decrease) content compared to control HK. Control HK had a higher melting temperature (T-m = 61.9 degrees C) than from dehydrated (T-m = 54.2 degrees C) frogs when thermostability was tested using differential scanning fluorimetry. In silico phosphorylation of a Xenopus HK caused alterations in active site binding, corroborating phosphorylation as the probable mechanism for kinetic regulation. Physiological consequences of dehydration-induced HK phosphorylation appear to facilitate glycolytic metabolism in hypoxic situations. Augmented HK function increases the ability of Xenopus to overcome compromised oxidative phosphorylation associated with ischemia during dehydration.
机译：非洲爪蟾忍受大量脱水，由于血流受损，可能会导致缺氧应激。组织可能会增加对厌氧糖酵解产生能量的依赖性，因此调节己糖激酶（HK）很重要。我们研究了来自对照青蛙和脱水青蛙（损失了30％的体内总水分）肌肉的纯化HK的酶学性质和磷酸化状态。生物信息学工具也被用于分析HK磷酸化在计算机中的结构意义。与对照HK相比，脱水青蛙肌肉的HK表现出葡萄糖的V-max（3.4倍）和K-m显着更高（2.4倍），但ATP的Km不变。与对照HK相比，来自脱水青蛙的HK还显示出更高的磷酸丝氨酸含量（增加了20％）和更低的磷酸苏氨酸含量（降低了22％）。当使用差示扫描荧光法测试热稳定性时，对照HK的融解温度（T-m = 61.9摄氏度）比脱水青蛙（T-m = 54.2摄氏度）更高。非洲爪蟾HK的计算机磷酸化导致活性位点结合的改变，证实了磷酸化是动力学调节的可能机制。缺氧情况下，脱水诱导的HK磷酸化的生理后果似乎促进糖酵解代谢。增强的HK功能增加了非洲爪蟾克服脱水过程中与局部缺血相关的受损的氧化磷酸化的能力。

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