Literature summary for 5.6.1.3 extracted from

Chang, Q.; Nitta, R.; Inoue, S.; Hirokawa, N.
Structural basis for the ATP-induced isomerization of kinesin (2013), J. Mol. Biol., 425, 1869-1880.

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	microtubule-stimulated ATPase activity of wild-type monomeric KIF4	Mus musculus

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of a KIF4 motor domain construct, KIF4-344 comprising residues 1-344 of KIF4 followed by a His7-tag in Escherichia coli strain BL21(DE3)	Mus musculus

Crystallization (Commentary)

Crystallization (Comment)	Organism
His7-tagged KIF4 motor domain construct KIF4-344 in complex with AMPPNP, sitting drop vapour diffusion method, mixing of 15 mg/ml protein in 10 mM TrisHCl, pH 8.0, 100 mM NaCl, and 1 mM DTT with a reservoir solution containing 28% w/v PEG 4000, 100 mM Tris-HCl, pH 8.5, and 200 mM sodium acetate, in the presence of 5 mM AMPPNP, X-ray diffraction strcuture determination and analysis at 1.71 A resolution, molecular replacement and modeling	Mus musculus

Crystallization (Comment)

Organism

His7-tagged KIF4 motor domain construct KIF4-344 in complex with AMPPNP, sitting drop vapour diffusion method, mixing of 15 mg/ml protein in 10 mM TrisHCl, pH 8.0, 100 mM NaCl, and 1 mM DTT with a reservoir solution containing 28% w/v PEG 4000, 100 mM Tris-HCl, pH 8.5, and 200 mM sodium acetate, in the presence of 5 mM AMPPNP, X-ray diffraction strcuture determination and analysis at 1.71 A resolution, molecular replacement and modeling

Mus musculus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics, and steady-state ATPase activities of three types of chimeric construct, overview	Mus musculus
0.00634	-	ATP	wild-type KIF4 enzyme, pH and temperature not specified in the publication	Mus musculus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + H2O + a kinesin associated with a microtubule at position n	Mus musculus	-	ADP + phosphate + a kinesin associated with a microtubule at position n+1 (toward the plus end)	-	?
additional information	Mus musculus	KIF4 specifically binds to the microtubule, enzyme interaction with tubulin, the KIF4-specific Q248 and K249 form possible hydrogen bonds with Y108 of helix H3' of alpha-tubulin. Microtubule filament binding by enzyme KIF4, around the center line of the microtubule protofilament, the helix-alpha4-mediated microtubule-binding site is located and fitted into the intra-tubulin dimer groove	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	P33174	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His7-tagged KIF4 motor domain construct KIF4-344 from Escherichia coli strain BL21(DE3) by nickel affinity and cation exchange chromatography, followed gel filtration and dialysis	Mus musculus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + H2O + a kinesin associated with a microtubule at position n	-	Mus musculus	ADP + phosphate + a kinesin associated with a microtubule at position n+1 (toward the plus end)	-	?
ATP + H2O + a kinesin associated with a microtubule at position n	nucleotide-induced conformational change catalytic method	Mus musculus	ADP + phosphate + a kinesin associated with a microtubule at position n+1 (toward the plus end)	-	?
additional information	KIF4 specifically binds to the microtubule, enzyme interaction with tubulin, the KIF4-specific Q248 and K249 form possible hydrogen bonds with Y108 of helix H3' of alpha-tubulin. Microtubule filament binding by enzyme KIF4, around the center line of the microtubule protofilament, the helix-alpha4-mediated microtubule-binding site is located and fitted into the intra-tubulin dimer groove	Mus musculus	?	-	?

Synonyms

Synonyms	Comment	Organism
KIF4	-	Mus musculus
KIF4 ATPase	-	Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
42.1	-	ATP	wild-type KIF4 enzyme, pH and temperature not specified in the publication	Mus musculus

General Information

General Information	Comment	Organism
evolution	the enzyme KIF4 belongs to the kinesin superfamily	Mus musculus
additional information	tubulin and ATP binding structure analysis, sequential docking of the neck linker induced by ATP binding (isomerization), modeling. Backdoor formation stabilizes the two switch regions and induces full docking of the neck linker through a conformational change to switch II, affecting the conformation of loops L11-alpha4-L12-alpha5-L13 in switch II. Overall architecture of the KIF4 motor domain and structure-function relationship, overview	Mus musculus
physiological function	kinesin superfamily proteins are microtubule-based molecular motors driven by the energy derived from the hydrolysis of ATP. The ATP binding step is crucial both for the power stroke to produce motility and for the inter-domain regulation of ATPase activity to guarantee the processive movement of dimeric kinesin superfamily proteins. The binding of KIF4 to the microtubule may affect the interaction within the tubulin dimer by increasing or decreasing the curvature of the intra-tubulin dimer interface with some twist, molecular mechanism by which KIF4 inhibits microtubule dynamics	Mus musculus