The linker between the dimerization and catalytic domains of the CheA histidine kinase propagates changes in structure and dynamics that are important for enzymatic activity.
The histidine kinase, CheA, couples environmental stimuli to changes in bacterial swimming behavior, converting a sensory signal to a chemical signal in the cytosol via autophosphorylation. The kinase activity is regulated in the platform of chemotaxis signaling complexes formed by CheW, chemoreceptors, and the regulatory domain of CheA. Our previous computational and mutational studies have revealed that two interdomain linkers play important roles in CheA's enzymatic activity. Of the two linkers, one that connects the dimerization and ATP binding domains is essential for both basal autophosphorylation and activation of the kinase. However, the mechanistic role of this linker remains unclear, given that it is far from the autophosphorylation reaction center (the ATP binding site). Here we investigate how this interdomain linker is coupled to CheA's enzymatic activity. Using modern nuclear magnetic resonance (NMR) techniques, we find that by interacting with the catalytic domain, the interdomain linker initiates long-range structural and dynamic changes directed toward the catalytic center of the autophosphorylation reaction. Subsequent biochemical assays define the functional relevance of these NMR-based observations. These findings extend our understanding of the chemotaxis signal transduction pathway.
Department of Chemistry and Biochemistry, University of California , Santa Barbara, California 93106-9510, United States.
Xiqing
X
Wang
Pramodh
P
Vallurupalli
Anh
A
Vu
Kwangwoon
K
Lee
Sheng
S
Sun
Wen-Ju
WJ
Bai
Chun
C
Wu
Hongjun
H
Zhou
Joan-Emma
JE
Shea
Lewis E
LE
Kay
Frederick W
FW
Dahlquist
D001426:Bacterial Proteins / Q000737:chemistry*; D020134:Catalytic Domain; D004789:Enzyme Activation; D000071677:Histidine Kinase; D008958:Models, Molecular; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry*; D055503:Protein Multimerization; D020124:Thermotoga maritima / Q000378:metabolism*
D001426:Bacterial Proteins; D011494:Protein Kinases; D000071677:Histidine Kinase