Title
Comparative proteomics reveal distinct chaperone–client interactions in supporting bacterial acid resistance
Author
Shuai Zhang, Dan He, Yi Yang, Shixian Lin, Meng Zhang, Shizhong Dai, Peng R. Chen
Year
2016
Journal
PNAS
Abstract
HdeA and HdeB are essential chaperones for many Gram-negative enteric bacteria to survive acid stress. How they cooperate to protect a broad range of client proteins from acid denaturation while avoiding nonspecific binding remains elusive. Here we developed a comparative proteomic strategy combining genetically encoded releasable photocross-linking with 2D difference gel electrophoresis, which allows an unbiased side-by-side comparison of the entire client pools from these two acid-activated chaperones in Escherichia coli. Our results reveal the existence of client specificities between these chaperones, and this pH-regulated client specificity mechanism demonstrates their seemingly paradoxical specific but promiscuous client recognition. Our comparative proteomic strategy provides a powerful platform for profiling protein–protein interactions under harsh conditions and for elucidating the otherwise elusive mode of action within native cellular contexts.
Instrument
J-815
Keywords
Circular dichroism, Secondary structure, Chemical stability, Biochemistry