Structural Alternation in Heat Shock Proteins of Activated Macrophages.
Structural Alternation in Heat Shock Proteins of Activated Macrophages.
复制标题
活化巨噬细胞热激蛋白的结构改变
作者：
Zhang W;Wei Y;Zhang H;Liu J;Zong Z;Liu Z;Zhu S;Hou W;Chen Y;Deng H
巨噬细胞的炎症反应是一个在严格调控下有序而复杂的过程，伴随着形态和功能的急剧变化。据预测，蛋白质将在这些精细调控的过程中发生结构变化。然而，在炎症反应过程中巨噬细胞的结构蛋白质组的变化仍然很差的特点。在本研究中，我们应用有限的蛋白水解偶联质谱（LiP-MS），以确定脂多糖（LPS）激活的巨噬细胞的蛋白质组结构的变化。我们从230个蛋白质中鉴定了386个结构特异性蛋白水解指纹。利用基因本体（GO）生物过程富集，我们发现结构改变的蛋白质被富集到蛋白质折叠相关的术语中，其中HSP 60被列为变化最大的蛋白质。我们通过细胞热位移分析（CETSA）和天然CETSA验证了HSP 60的结构变化。结果表明，活化的巨噬细胞中HSP 60的热稳定性增强，并形成一种不含HSP 10的复合物。总之，我们表明，在原位结构系统生物学是一种有效的方法来表征蛋白质组结构的变化，并揭示伴侣蛋白的结构变化显着在巨噬细胞活化。
The inflammatory response of macrophages is an orderly and complex process under strict regulation accompanied by drastic changes in morphology and functions. It is predicted that proteins will undergo structural changes during these finely regulated processes. However, changes in structural proteome in macrophages during the inflammatory response remain poorly characterized. In the present study, we applied limited proteolysis coupled mass spectrometry (LiP-MS) to identify proteome-wide structural changes in lipopolysaccharide (LPS)-activated macrophages. We identified 386 structure-specific proteolytic fingerprints from 230 proteins. Using the Gene Ontology (GO) biological process enrichment, we discovered that proteins with altered structures were enriched into protein folding-related terms, in which HSP60 was ranked as the most changed protein. We verified the structural changes in HSP60 by using cellular thermal shift assay (CETSA) and native CETSA. Our results showed that the thermal stability of HSP60 was enhanced in activated macrophages and formed an HSP10-less complex. In conclusion, we demonstrate that in situ structural systems biology is an effective method to characterize proteomic structural changes and reveal that the structures of chaperone proteins vary significantly during macrophage activation.
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影响因子：
7.3
作者：
Canton M;Sánchez-Rodríguez R;Spera I;Venegas FC;Favia M;Viola A;Castegna A
通讯作者：
Castegna A
影响因子：
7.4
作者：
Thompson, A;Schäfer, J;Hamon, C
通讯作者：
Hamon, C
影响因子：
5.8
作者：
Walter, Wencke;Sanchez-Cabo, Fatima;Ricote, Mercedes
通讯作者：
Ricote, Mercedes
影响因子：
64.8
作者：
Xu, ZH;Horwich, AL;Sigler, PB
通讯作者：
Sigler, PB
影响因子：
4.8
作者：
Zhou, YQ;Chen, YQ;Wang, MH
通讯作者：
Wang, MH