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Updated: Jul 14, 2026
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Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
Published on: December 27, 2016
NMR structure of activated CheY
1Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA, 94720, USA.
Journal of Molecular Biology
|March 25, 2000
Summary
Researchers stabilized the active state of the CheY protein, crucial for bacterial chemotaxis, using beryllofluoride. This allowed detailed structural analysis, revealing key rearrangements for protein activation.
Area of Science:
- Biochemistry
- Structural Biology
- Microbiology
Background:
- CheY protein acts as a response regulator in bacterial chemotaxis.
- Phosphorylation triggers structural changes, converting CheY from inactive to active.
- The transient nature of phosphorylated CheY hinders detailed structural studies of its active form.
Purpose of the Study:
- To achieve persistent activation of Escherichia coli CheY.
- To determine the structure of the active CheY protein using NMR spectroscopy.
- To elucidate the structural mechanisms underlying CheY activation.
Main Methods:
- Persistent activation of CheY via complexation with beryllofluoride (BeF(3)(-)).
- Nuclear Magnetic Resonance (NMR) spectroscopy for structural determination.
- Analysis of structural changes and hydrogen bond formation.
Main Results:
- The structure of activated CheY was determined with high resolution (backbone r.m.s.d. of 0.58 Å).
- A key hydrogen bond formed between Thr87 and Asp57.BeF(3)(-) stabilized the active conformation.
- A coupled rearrangement of conserved residues (Thr87, Tyr106) and displacement of beta4/H4 were observed.
Conclusions:
- The study provides the first detailed structure of persistently activated CheY.
- Stabilization via hydrogen bonding and coupled residue rearrangement are critical for CheY activation.
- This mechanism of activation may be applicable to other receiver domains in signaling pathways.
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