Literature summary for 5.6.1.3 extracted from

Yokoyama, H.; Sawada, J.; Katoh, S.; Matsuno, K.; Ogo, N.; Ishikawa, Y.; Hashimoto, H.; Fujii, S.; Asai, A.
Structural basis of new allosteric inhibition in kinesin spindle protein Eg5 (2015), ACS Chem. Biol., 10, 1128-1136.

Application

Application	Comment	Organism
medicine	inhibition of mitotic spindles is a pharmaceutically validated strategy for cancer therapeutics, human Eg5 in addition to microtubules represents an attractive target molecule for novel clinical therapies in the treatment of human malignancies, kinesin spindle protein Eg5 is a target for anticancer therapies	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
gene KIF11, recombinant expression of C-terminally His6-tagged wild-type Eg51-369 and mutants in Escherichia coli BL21(DE3)pLysS	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant Eg5 motor domain, residues 1-369, in complex with inhibitor PVZB1194, sitting drop vapor diffusion method, at 4°C, mixing of 500 nl of 6 mg/ml protein in 50 mM PIPES-NaOH, pH 6.8, 250 mM NaCl, 2 mM MgCl2, 1 mM EGTA-NaOH, 1 mM TCEP-HCl, and 10% w/v sucrose, and inhibitor PVZB1194 (ratio 1:3 or 1:4 with enzyme) solution with 500 nl of the reservoir solution containing 19% w/v PEG 3350, 0.1 M MES-NaOH, pH 6.0, and 200 mM NaNO3, 1 week, followed by microseeding, X-ray diffraction structure determination and analysis at 2.8 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant Eg5 motor domain, residues 1-369, in complex with inhibitor PVZB1194, sitting drop vapor diffusion method, at 4°C, mixing of 500 nl of 6 mg/ml protein in 50 mM PIPES-NaOH, pH 6.8, 250 mM NaCl, 2 mM MgCl2, 1 mM EGTA-NaOH, 1 mM TCEP-HCl, and 10% w/v sucrose, and inhibitor PVZB1194 (ratio 1:3 or 1:4 with enzyme) solution with 500 nl of the reservoir solution containing 19% w/v PEG 3350, 0.1 M MES-NaOH, pH 6.0, and 200 mM NaNO3, 1 week, followed by microseeding, X-ray diffraction structure determination and analysis at 2.8 A resolution

Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	development of small molecule inhibitors of the enzyme Kif11 ATPase activity	Homo sapiens
PVZB1194	allosteric inhibitor, biphenyl-type inhibitor, binding structure of the inhibitor in complex with the Eg5 motor domain: inhibitor PVZB1194 binds to the alpha4/alpha6 allosteric pocket 15 A from the ATP-binding pocket, which differs from conventional allosteric inhibitors that bind to the allosteric L5/alpha2/alpha 3 pocket of Eg5. Binding of the inhibitor is involved in the neck-linker conformation and also causes conformational changes around the ATP-binding pocket through Tyr104 to affect the interaction of ATP with the pocket. Folding rearrangement of enzyme Eg5 induced by PVZB1194 in the absence of nucleotides and microtubules. Residue Tyr104 is involved in ATP-competitive inhibition by PVZB1194	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Homo sapiens

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	Homo sapiens	-	ADP + phosphate + a kinesin associated with a microtubule at position n+1 (toward the plus end)	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P52732	gene KIF11	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His6-tagged wild-type Eg51-369 and mutants from Escherichia coli BL21(DE3)pLysS by nickel affinity and cation exchange chromatography	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + H2O + a kinesin associated with a microtubule at position n	-	Homo sapiens	ADP + phosphate + a kinesin associated with a microtubule at position n+1 (toward the plus end)	-	?

Synonyms

Synonyms	Comment	Organism
Eg5	-	Homo sapiens
KIF11	-	Homo sapiens
kinesin spindle protein	-	Homo sapiens
kinesin-like protein	-	Homo sapiens

General Information

General Information	Comment	Organism
evolution	the enzyme is a member of the kinesin-5 family	Homo sapiens
malfunction	inhibition of Eg5 causes cell cycle arrest in mitosis with the irregular formation of monopolar spindles and subsequent apoptotic cell death	Homo sapiens
additional information	analysis of the molecular mechanism responsible for regulating the motor activity of kinesins, it plays an essential role in centrosome separation and bipolar mitotic spindle formation during the early stage of mitosis	Homo sapiens
physiological function	kinesin spindle protein, i.e. Eg5 or KIF11, is a mitotic spindle motor protein	Homo sapiens