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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Thermal disaggregation of type B yeast hexokinase by indole derivatives: A mechanistic study
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Thermal disaggregation of type B yeast hexokinase by indole derivatives: A mechanistic study

机译：吲哚衍生物热分解B型酵母己糖激酶的机理研究

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摘要

Protein aggregation is a pathological hallmark of several human disorders, and a central problem in biotechnology, occurring during purification, sterilization, shipping and storage of protein structures. The process is a very complex one, characterized with a remarkable polymorphism of aggregates, including soluble amyloid oligomers, amyloid fibrils and amorphous species. While amyloid structure formation has been extensively investigated during the recent years, amorphous aggregation is still not well characterized. Use of small molecules that affect this process could be informative in this regard. In order to explore the inhibiting effect of small molecules on the amorphous aggregate formation, yeast hexokinase-B, a key enzyme in metabolism, has been chosen for the present study. Thermal aggregation of the enzyme was investigated in 50. mM phosphate buffer, pH 7 at 55 °C and the extent of aggregation was measured by monitoring the increase in absorbance at 350. nm versus time. Possible anti-aggregation effects of a variety of non-specific ligands including indole, tryptophan, carbinol, and indomethacin were explored. Turbidity of the protein solutions was found to be diminished by the presence of these small molecules in the above conditions, with the highest effects being exerted by indomethacin. Dynamic light scattering and HPLC confirmed that indomethacin had the highest anti-aggregation effect. These observations, taken together, suggest that the indole ring is likely to play an important role in aggregation inhibition.
机译：蛋白质聚集是几种人类疾病的病理特征，也是生物技术中的一个核心问题，在蛋白质结构的纯化，灭菌，运输和存储过程中会发生。该过程是一个非常复杂的过程，其特征是聚集体具有明显的多态性，包括可溶性淀粉样蛋白低聚物，淀粉样蛋白原纤维和无定形物种。尽管近年来对淀粉样蛋白结构的形成进行了广泛的研究，但非晶聚集仍然没有得到很好的表征。在这方面，使用影响该过程的小分子可能会有所帮助。为了探索小分子对无定形聚集体形成的抑制作用，本研究选择了酵母己糖激酶-B（一种代谢中的关键酶）。在55°C的pH为7的50. mM磷酸盐缓冲液中研究了酶的热聚集，并通过监测350. nm处的吸光度随时间的增加来测量聚集程度。探索了各种非特异性配体（包括吲哚，色氨酸，甲醇和吲哚美辛）的可能抗聚集作用。在上述条件下，这些小分子的存在降低了蛋白质溶液的浊度，消炎痛的作用最大。动态光散射和HPLC证实消炎痛具有最高的抗聚集作用。这些观察结果加在一起，表明吲哚环可能在聚集抑制中起重要作用。

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