Literature summary for 2.7.13.3 extracted from

Greenswag, A.R.; Muok, A.; Li, X.; Crane, B.R.
Conformational transitions that enable histidine kinase autophosphorylation and receptor array integration (2015), J. Mol. Biol., 427, 3890-3907.

Crystallization (Commentary)

Crystallization (Comment)	Organism
P3P4 domain structure of Thermotoga maritima CheA, vapor diffusion method, mixing of 0.001 ml of 0.7 mM protein in 50 mM HEPES, pH 7.5, 150 mM NaCl, and 2 mM DTT, with 0.001 ml of reservoir solution containing 0.5 M ammonium sulfate, 0.1 M sodium citrate tribasic dihydrate, pH 5.6, and 1.0 M lithium sulfate monohydrate, 4°C, X-ray small angle diffraction structure determination and analysis at 3.0 A resolution, molecular replacement and modelling	Thermotoga maritima

Crystallization (Comment)

Organism

P3P4 domain structure of Thermotoga maritima CheA, vapor diffusion method, mixing of 0.001 ml of 0.7 mM protein in 50 mM HEPES, pH 7.5, 150 mM NaCl, and 2 mM DTT, with 0.001 ml of reservoir solution containing 0.5 M ammonium sulfate, 0.1 M sodium citrate tribasic dihydrate, pH 5.6, and 1.0 M lithium sulfate monohydrate, 4°C, X-ray small angle diffraction structure determination and analysis at 3.0 A resolution, molecular replacement and modelling

Thermotoga maritima

Protein Variants

Protein Variants	Comment	Organism
D449A	site-directed mutagenesis, the mutation in the ATP-binding pocket prevents nucleotide binding	Thermotoga maritima
H405Y	site-directed mutagenesis, the mutation abrogates the kinase activity	Thermotoga maritima
H45K	site-directed mutagenesis, a CheAFL variant that lacks the substrate His	Thermotoga maritima
H45K/H405Y	site-directed mutagenesis, the mutant shows reduced kinase activity compared to the wild-type	Thermotoga maritima
H45K/S492C	site-directed mutagenesis	Thermotoga maritima
additional information	disulfide cross-linking of mutant enzymes, overview	Thermotoga maritima
S492C	site-directed mutagenesis	Thermotoga maritima

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + protein L-histidine	Thermotoga maritima	-	ADP + protein N-phospho-L-histidine	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
phosphoprotein	histidine kinase performs autophosphorylation	Thermotoga maritima

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + protein L-histidine	-	Thermotoga maritima	ADP + protein N-phospho-L-histidine	-	?
additional information	histidine kinase performs autophosphorylation. The His-containing substrate domain (P1) is sequestered by interactions that depend upon P1 of the adjacent subunit. Non-hydrolyzable ATP analogues (but not ATP or ADP) release P1 from the protein core (domains P3P4P5) and increase its mobility. Autophosphorylation is possible only when the subunit with a functional P4 domain trans phosphorylates a functional P1 domain of the opposing subunit	Thermotoga maritima	?	-	?

Synonyms

Synonyms	Comment	Organism
CheA	-	Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Thermotoga maritima

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Thermotoga maritima

Cofactor

Cofactor	Comment	Organism	Structure
ATP	residue Ser492 resides on the ATP lid, near to the Mg2+ ion that coordinates the gamma-phosphate of ATP in the P4 active site	Thermotoga maritima
additional information	non-hydrolyzable ATP analogues (but not ATP or ADP) release P1 from the protein core (domains P3P4P5) and increase its mobility	Thermotoga maritima

General Information

General Information	Comment	Organism
evolution	CheA differs from sensor histidine kinases in several ways: CheA does not contain a transmembrane domain, relying instead on P5 and CheW for interaction with transmembrane components. It has the phosphorylatable His residue on a separate domain (P1) instead of the dimerization domain (P3), and it utilizes a separate docking domain (P2) for CheY. P2 is not necessary for phosphotransfer to the response regulator CheY per se but variants lacking the P2 domain (DELTAP2) exhibit a reduced phosphotransfer rate relative to full-length CheA (CheAFL) and support a lower extent of chemotaxis. The linkers between the CheA domains play important roles in CheA activity	Thermotoga maritima
physiological function	CheA differs from sensor histidine kinases in several ways: CheA does not contain a transmembrane domain, relying instead on P5 and CheW for interaction with transmembrane components. It has the phosphorylatable His residue on a separate domain (P1) instead of the dimerization domain (P3), and it utilizes a separate docking domain (P2) for CheY. P2 is not necessary for phosphotransfer to the response regulator CheY per se but variants lacking the P2 domain (DELTAP2) exhibit a reduced phosphotransfer rate relative to full-length CheA (CheAFL) and support a lower extent of chemotaxis. The linkers between the CheA domains play important roles in CheA activity	Thermotoga maritima