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Effect of Thaw Temperatures in Reducing Enzyme Activity in Human Thyroid Tissues

机译：解冻温度对降低人甲状腺组织酶活性的影响

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An identified impediment to the advancement of science in the field of proteomics is the deterioration of proteins in tissue upon removal of the tissue from its natural state. To reduce this degradation, human tissues are frozen and stored in either liquid nitrogen or - 80℃ environments. It is believed that frozen tissue in ultralow temperatures preserves proteins against enzyme degradation. Various molecular, biophysical, and biochemical analytical studies require that frozen tissues be thawed before being used for analyses. Depending on downstream analyses, tissues are thawed at different temperatures （37℃, room temperature or 4℃）. However, there is very little literature that describes the effects of different thaw temperatures on enzymatic inactivation in tissue lysates. We investigated the effects of preprocessing variable thaw temperature on postprocessed lysates using tyrosine phosphatase and phosphatase and tensin homolog activity assays. In our study we examined the thawing of frozen human thyroid tissues at the traditional temperatures of 4℃ (on ice), 37℃ (in an oven), and the novel temperature of 95℃ (using Stabilizor T1~(?)). The tissue lysates were processed without the addition of enzymatic inhibitors. Our results showed that in benign, malignant, and diseased tissues, high temperature thawing is effective in reducing enzymatic activity. In normal tissue, the reduction is dependent on individual enzymes. This suggests that if tissue lysates are to be obtained from frozen tissues without the addition of inhibitors, high temperature thawing might have marked improvement in downstream non-enzymatic analyses of diseased and neoplastic tissues.
机译：蛋白质组学领域科学发展的一个公认障碍是，从自然状态中取出组织后组织中蛋白质的降解。为了减少这种降解，将人体组织冷冻并储存在液氮或-80℃的环境中。可以相信，在超低温下的冷冻组织可以保护蛋白质免受酶降解。各种分子，生物物理和生化分析研究要求将冷冻的组织融化后再用于分析。根据下游分析，在不同温度（37℃，室温或4℃）下解冻组织。但是，很少有文献描述不同解冻温度对组织裂解物中酶促失活的影响。我们使用酪氨酸磷酸酶，磷酸酶和张力蛋白同源物活性测定法研究了可变解冻温度对后处理裂解物的影响。在我们的研究中，我们研究了在传统温度4℃（冰上），37℃（在烤箱中）和95℃的新温度（使用Stabilizor T1〜（？））下冷冻的人体甲状腺组织的解冻情况。在不添加酶抑制剂的情况下处理组织裂解物。我们的结果表明，在良性，恶性和患病组织中，高温解冻可有效降低酶的活性。在正常组织中，减少取决于单个酶。这表明，如果要在不添加抑制剂的情况下从冷冻组织中获得组织裂解物，则高温融化可能在患病和赘生性组织的下游非酶分析中有显着改善。

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