Structural Alternation in Heat Shock Proteins of Activated Macrophages

Cells

Zhaoyun Zong¹  Wenxuan Hou¹  Ying Wei¹  Huaijin Zhang¹  Haiteng Deng¹  Wenhao Zhang¹  Jing Liu¹  Yuling Chen¹  Songbiao Zhu¹  Zongyuan Liu¹ 

[1] MOE Key Laboratory of Bioinformatics, Center for Synthetic and Systematic Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China;

关键词: structural proteomics;    RAW264.7;    LPS;    Lip-MS;    HSP60;   

DOI  :  10.3390/cells10123507

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown