Literature summary for 2.7.11.21 extracted from

Kothe, M.; Kohls, D.; Low, S.; Coli, R.; Cheng, A.C.; Jacques, S.L.; Johnson, T.L.; Lewis, C.; Loh, C.; Nonomiya, J.; Sheils, A.L.; Verdries, K.A.; Wynn, T.A.; Kuhn, C.; Ding, Y.H.
Structure of the catalytic domain of human polo-like kinase 1 (2007), Biochemistry, 46, 5960-5971.

Search for more literature for 2.7.11.21

Application

Application	Comment	Organism
medicine	Plk1 is a target for the development of anticancer therapeutics	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in sf21 cells	Homo sapiens
into pFastBac1 for expression in Spodoptera frugiperda insect cells	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structures of the T210V mutant of the kinase domain of human Plk1 complexed with adenylylimidodiphosphate or the pyrrolo-pyrazole inhibitor PHA-680626 are determined at 2.4 and 2.1 A resolution, respectively	Homo sapiens
wild-type, to ca. 3 A resolution, T210V mutant complexed with the nonhydrolyzable ATP analogue AMP-PNP or the pyrrolo-pyrazole inhibitor PHA-680626 at 2.4 and 2.1 A resolution, respectively, by the hanging drop method, typical kinase fold with the unphosphorylated (mutant T210V) activation loop in an extended conformation, stabilized by a crystal contact, presence of a phenylalanine at the bottom of the ATP pocket, combined with a cysteine in the roof of the binding site, a pocket created by Leu132 in the hinge region, and a cluster of positively charged residues in the solvent-exposed area outside of the adenine pocket adjacent to the hinge region	Homo sapiens

Crystallization (Comment)

Organism

crystal structures of the T210V mutant of the kinase domain of human Plk1 complexed with adenylylimidodiphosphate or the pyrrolo-pyrazole inhibitor PHA-680626 are determined at 2.4 and 2.1 A resolution, respectively

Homo sapiens

wild-type, to ca. 3 A resolution, T210V mutant complexed with the nonhydrolyzable ATP analogue AMP-PNP or the pyrrolo-pyrazole inhibitor PHA-680626 at 2.4 and 2.1 A resolution, respectively, by the hanging drop method, typical kinase fold with the unphosphorylated (mutant T210V) activation loop in an extended conformation, stabilized by a crystal contact, presence of a phenylalanine at the bottom of the ATP pocket, combined with a cysteine in the roof of the binding site, a pocket created by Leu132 in the hinge region, and a cluster of positively charged residues in the solvent-exposed area outside of the adenine pocket adjacent to the hinge region

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
T210D	mutant	Homo sapiens
T210D	expression at low levels, no crystals obtained	Homo sapiens
T210V	mutant	Homo sapiens
T210V	crystals obtained	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
AMPPNP	-	Homo sapiens
PHA-680626	selective against Plk1	Homo sapiens
scytonemin	-	Homo sapiens
Wortmannin	-	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0006	-	Cdc25C	mutant T210D	Homo sapiens
0.0006	-	Cdc25C	apparent Km, T210D mutant, varied substrate Cdc25c, constant substrate ATP	Homo sapiens
0.00066	-	Cdc25C	wild-type	Homo sapiens
0.00066	-	Cdc25C	apparent Km, wild-type enzyme, varied substrate Cdc25c, constant substrate ATP	Homo sapiens
0.0008	-	Cdc25C	mutant T210V	Homo sapiens
0.0008	-	Cdc25C	apparent Km, T210V mutant, varied substrate Cdc25c, constant substrate ATP	Homo sapiens
0.003	-	ATP	mutant T210V	Homo sapiens
0.003	-	ATP	apparent Km, T210V mutant, varied substrate ATP, constant substrate Cdc25c	Homo sapiens
0.0032	-	ATP	wild-type	Homo sapiens
0.0032	-	ATP	apparent Km, wild-type enzyme, varied substrate ATP, constant substrate Cdc25c	Homo sapiens
0.004	-	ATP	mutant T210D	Homo sapiens
0.004	-	ATP	apparent Km, T210D mutant, varied substrate ATP, constant substrate Cdc25c	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + protein	Homo sapiens	-	ADP + phosphoprotein	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-
Homo sapiens	P53350	-	-

Purification (Commentary)

Purification (Comment)	Organism
on nickel affinity column and by gel filtration	Homo sapiens
using a HiTrap nickel chelating affinity column and a gel filtration column, the His-tag is removed by PreScission protease treatment	Homo sapiens

Storage Stability

Storage Stability	Organism
-80°C, 50 mM HEPES buffer, pH 7.5, 5 mM TCEP	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + Cdc25C	-	Homo sapiens	ADP + phosphorylated Cdc25C	-	?
ATP + Cdc25C	substrate in activity assay	Homo sapiens	ADP + phosphorylated Cdc25C	-	?
ATP + protein	-	Homo sapiens	ADP + phosphoprotein	-	?

Synonyms

Synonyms	Comment	Organism
Plk1	-	Homo sapiens
Polo-like kinase 1	-	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.0019	-	ATP	mutant T210V	Homo sapiens
0.0019	-	ATP	T210V mutant, varied substrate Cdc25c, constant substrate ATP	Homo sapiens
0.0041	-	ATP	wild-type	Homo sapiens
0.0041	-	ATP	wild-type enzyme, varied substrate ATP, constant substrate Cdc25c	Homo sapiens
0.0049	-	Cdc25C	mutant T210V	Homo sapiens
0.0049	-	Cdc25C	T210V mutant, varied substrate ATP, constant substrate Cdc25c	Homo sapiens
0.0094	-	Cdc25C	wild-type	Homo sapiens
0.0094	-	Cdc25C	wild-type enzyme, varied substrate Cdc25c, constant substrate ATP	Homo sapiens
0.019	-	ATP	mutant T210D	Homo sapiens
0.019	-	ATP	T210D mutant, varied substrate ATP, constant substrate Cdc25c	Homo sapiens
0.0376	-	Cdc25C	T210D mutant, varied substrate Cdc25c, constant substrate ATP	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	activity assay	Homo sapiens

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.000024	-	-	Homo sapiens	Wortmannin
0.00034	-	mutant T210D	Homo sapiens	PHA-680626
0.00044	-	mutant T210V	Homo sapiens	PHA-680626
0.00053	-	-	Homo sapiens	PHA-680626
0.00053	-	wild-type	Homo sapiens	PHA-680626
0.002	-	-	Homo sapiens	scytonemin
0.14	-	mutant T210D	Homo sapiens	AMPPNP
0.155	-	mutant T210V	Homo sapiens	AMPPNP
0.166	-	wild-type	Homo sapiens	AMPPNP