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Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
Published on: September 28, 2018
Antagonism between binding site affinity and conformational dynamics tunes alternative cis-interactions within Shp2
Jie Sun1, Shaoying Lu, Mingxing Ouyang
1Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Nature Communications
|June 25, 2013
Summary
Protein conformation, crucial for function, is influenced by evolutionary dynamics. Cis-interactions in Shp2 protein tyrosine phosphatase are tuned by Grb2, revealing sequence and position interplay for tunable protein conformations.
Area of Science:
- Molecular Biology
- Biochemistry
- Evolutionary Biology
Background:
- Protein function is intrinsically linked to its three-dimensional conformation.
- Modular proteins, composed of distinct domains, exhibit complex conformational dynamics.
- The evolutionary mechanisms shaping these protein conformations are not fully understood.
Purpose of the Study:
- To investigate the evolutionary dynamics governing the conformation of modular proteins.
- To elucidate how cis-interactions within the protein tyrosine phosphatase Shp2 are regulated.
- To understand the role of adaptor protein Grb2 in modulating Shp2 conformation.
Main Methods:
- Analysis of cis-interactions between C-terminal phosphotyrosines and the SH2 domain of Shp2.
- Investigating the influence of adaptor protein Grb2 on these interactions.
- Utilizing sequence and positional analysis of phosphotyrosines (pY542 and pY580).
- Performing sequence swapping experiments to assess conformational changes.
Main Results:
- Cis-interactions in Shp2 are tunable by the adaptor protein Grb2.
- The competitiveness of phosphotyrosines pY542 and pY580 is determined by an antagonistic interplay of amino acid sequence and position.
- pY580 exhibits an advantage due to favorable positioning despite an adverse sequence context.
- Swapping sequences of pY542 and pY580 leads to dominant cis-interaction and inhibits Grb2 trans-regulation.
Conclusions:
- An antagonistic combination of sequence and position acts as a fundamental design principle for tunable protein conformations.
- This mechanism provides insights into the evolutionary adaptation of modular protein structures.
- Understanding these principles is key to deciphering protein function and regulation.
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