Aggregation of granulocyte colony stimulating factor under physiological conditions: Characterization and thermodynamic inhibition
Aggregation of granulocyte colony stimulating factor under physiological conditions: Characterization and thermodynamic inhibition
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DOI：
10.1021/bi012006m
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发表时间：
2002-05-21
期刊：
影响因子：
2.9
通讯作者：
Carpenter, JF
中科院分区：
文献类型：
--
作者：
Krishnan, S;Chi, EY;Carpenter, JF
我们研究了重组人粒细胞集落刺激因子（rhGCSF）的聚集，rhGCSF是一种在pH值6.9和37度下快速聚集和沉淀的蛋白质。我们观察到，天然单体rhGCSF在生理条件下可逆地形成二聚体，并且这种二聚体物种不参与不可逆聚集过程。蔗糖，一种热力学稳定剂，抑制rhGCSF的聚集。我们假设蔗糖通过降低结构扩展物种的浓度而起作用，这与优先排斥有利于原生状态集合中大多数紧凑物种的假设相一致。展开曲线的热力学稳定性数据和氢-氘交换实验结果支持上述假设。因此，在生理条件下使用热力学稳定剂稳定蛋白质的天然状态的策略，特别是与天然状态具有高亲和力的配体结合，有望防止在这种无干扰的溶液条件下发生蛋白质聚集。
We have investigated the aggregation of recombinant human granulocyte colony stimulating factor (rhGCSF), a protein that rapidly aggregates and precipitates at pH 6.9 and 37 degreesC. We observed that native monomeric rhGCSF reversibly forms a dimer under physiological conditions and that this dimeric species does not participate in the irreversible aggregation process. Sucrose, a thermodynamic stabilizer, inhibits the aggregation of rhGCSF. We postulate that sucrose acts by reducing the concentration of structurally expanded species, consistent with the hypothesis that preferential exclusion favors most compact species in the native state ensemble. Thermodynamic stability data from unfolding curves and hydrogen-deuterium exchange experimental results support the above hypothesis. Thus, the strategy of stabilizing the native state of the protein under physiological conditions using thermodynamic stabilizers, especially ligands binding with high affinity to the native state, is expected to protect against protein aggregation occurring under such nonperturbing solution conditions.