Activation of rat-liver microsomal glucose 6-phosphatase, inorganic pyrophosphatase and inorganic pyrophosphate-glucose phosphotransferase by hydroxyl ion

Marjorie R. Stetten, Foster F. Burnett
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引用次数: 51

Abstract

  • 1.

    1. OH- has been shown to produce a 2- to 3-fold increase in the apparent activity in vitro of rat-liver microsomal glucose 6-phosphatase (EC 3.1.3.9) and the related enzymatic activities, inorganic pyrophosphatase, inorganic pyrophosphate-glucose phosphotransferase and adenosine-5′-triphosphate-glucose phosphotransferase.

  • 2.

    2. Optimal activation is achieved by pre-treatment of liver microsomes with ammonium or amino acid buffers at pH 9.5–9.8. Inactivation is observed above pH 10.

  • 3.

    3. This activation is as great or greater than that produced by deoxycholate or Triton X-100 treatment and the thermal instability introduced by these reagents is avoided.

  • 4.

    4. Alteration of the microsomal membranes by treatment with base is accompanied by a decrease in turbidity but does not result in a solubilization of the enzyme. The essential protein-lipid binding is apparently retained and the enzyme, so treated, is relatively stable at 30°.

  • 5.

    5. NH4OH pretreatment of microsomes results in a lowering of the apparent Michaelis constant for glucose 6-phosphatase, similar to that caused by digitonin, deoxycholate and Triton X-100.

羟基离子对大鼠肝微粒体葡萄糖6-磷酸酶、无机焦磷酸酶和无机焦磷酸酶-葡萄糖磷酸转移酶的激活作用
1.1. OH-已被证明能使大鼠肝微粒体葡萄糖6-磷酸酶(EC 3.1.3.9)的体外表观活性以及相关酶,无机焦磷酸酶、无机焦磷酸-葡萄糖磷酸转移酶和腺苷-5 ' -三磷酸-葡萄糖磷酸转移酶的活性增加2- 3倍。在pH 9.5-9.8的条件下,用铵或氨基酸缓冲液预处理肝微粒体,可达到最佳活化效果。在pH 10.3.3以上观察到失活。这种活化与脱氧胆酸盐或Triton X-100处理产生的活化一样大或更大,并且避免了这些试剂引入的热不稳定性。用碱处理微粒体膜的改变伴随着浊度的减少,但不导致酶的增溶。必需的蛋白-脂结合显然被保留了下来,这样处理的酶在30°0.5.5下是相对稳定的。对微粒体进行NH4OH预处理后,葡萄糖6-磷酸酶表观Michaelis常数降低,与洋地黄苷、脱氧胆酸盐和Triton X-100的作用类似。
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