Visualizing an Ultra-Weak Protein-Protein Interaction in Phosphorylation Signaling

Qiong Xing, Peng Huang, Ju Yang, Jian Qiang Sun, Zhou Gong, Xu Dong, Da Chuan Guo, Shao Min Chen, Yu Hong Yang, Yan Wang, Ming Hui Yang, Ming Yi, Yi Ming Ding, Mai Li Liu, Wei Ping Zhang, Chun Tang

Research output: Contribution to journalArticlepeer-review

23 Scopus citations


Proteins interact with each other to fulfill their functions. The importance of weak protein-protein interactions has been increasingly recognized. However, owing to technical difficulties, ultra-weak interactions remain to be characterized. Phosphorylation can take place via a KD≈25 mM interaction between two bacterial enzymes. Using paramagnetic NMR spectroscopy and with the introduction of a novel GdIII-based probe, we determined the structure of the resulting complex to atomic resolution. The structure accounts for the mechanism of phosphoryl transfer between the two enzymes and demonstrates the physical basis for their ultra-weak interaction. Further, molecular dynamics (MD) simulations suggest that the complex has a lifetime in the micro- to millisecond regimen. Hence such interaction is termed a fleeting interaction. From mathematical modeling, we propose that an ultra-weak fleeting interaction enables rapid flux of phosphoryl signal, providing a high effective protein concentration. Phosphorylation signaling takes place between two bacterial enzymes EI and EIIAGlc, which have a binding affinity of only 25 mM (see picture). The structure of the ultra-weak fleeting complex was determined to atomic resolution by a novel paramagnetic NMR technique, and it shows that electrostatic repulsion largely accounts for the low affinity between the two proteins.

Original languageEnglish (US)
Pages (from-to)11501-11505
Number of pages5
JournalAngewandte Chemie - International Edition
Issue number43
StatePublished - Oct 1 2014
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


  • NMR spectroscopy
  • paramagnetic relaxation enhancement
  • protein-protein interactions
  • proteins
  • signal transduction


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