Literature summary for 3.6.4.13 extracted from

Iwatani-Yoshihara, M.; Ito, M.; Klein, M.G.; Yamamoto, T.; Yonemori, K.; Tanaka, T.; Miwa, M.; Morishita, D.; Endo, S.; Tjhen, R.; Qin, L.; Nakanishi, A.; Maezaki, H.; Kawamoto, T.
Discovery of allosteric inhibitors targeting the spliceosomal RNA helicase Brr2 (2017), J. Med. Chem., 60, 5759-5771 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His6-tagged enzyme in Spodooptera frugiperda Sf9 cells	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged enzyme in complex with inhibitors 6-benzyl-3-[(2R)-2-(3-fluoropyridin-2-yl)-6-methyl-3,4-dihydro-2H-1-benzopyran-7-yl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione, 6-benzyl-3-[3-(benzyloxy)phenyl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione, and 3-[5-[(Z)-(2-cyclohexyl-5-imino-7-oxo-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid, X-ray diffraction structure determination and analysis at 2.75 A, 2.95 A, and 2.80 A resolution, respectivly	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant detagged enzyme in complex with inhibitors 6-benzyl-3-[(2R)-2-(3-fluoropyridin-2-yl)-6-methyl-3,4-dihydro-2H-1-benzopyran-7-yl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione, 6-benzyl-3-[3-(benzyloxy)phenyl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione, and 3-[5-[(Z)-(2-cyclohexyl-5-imino-7-oxo-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid, X-ray diffraction structure determination and analysis at 2.75 A, 2.95 A, and 2.80 A resolution, respectivly

Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
(6Z)-6-[[5-(3-chlorophenyl)furan-2-yl]methylidene]-5-imino-2-propyl-5,6-dihydro-7H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-7-one	-	Homo sapiens
3-[5-[(Z)-(2-cyclohexyl-5-imino-7-oxo-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid	binds an RNA-binding site inside the N-terminal cassette, mechanism of action, enzyme binding structure, overview	Homo sapiens
3-[5-[(Z)-(5-imino-7-oxo-2-propyl-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid	-	Homo sapiens
6-benzyl-3-[(2R)-2-(3-fluoropyridin-2-yl)-6-methyl-3,4-dihydro-2H-1-benzopyran-7-yl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione	binds to an unexpected allosteric site between the C-terminal and the N-terminal helicase cassettes, enzyme binding structure, overview	Homo sapiens
6-benzyl-3-[3-(benzyloxy)phenyl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione	enzyme binding structure, overview	Homo sapiens
additional information	selectivity profiling indicated the allosteric inhibitor 6-benzyl-3-[(2R)-2-(3-fluoropyridin-2-yl)-6-methyl-3,4-dihydro-2H-1-benzopyran-7-yl]-4,6-dihydropyrido[4,3-d]pyrimidine-2,7(1H,3H)-dione is more Brr2-selective than the RNA site binder 3-[5-[(Z)-(2-cyclohexyl-5-imino-7-oxo-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid	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	Homo sapiens	-	ADP + phosphate	-	?
additional information	Homo sapiens	enzyme Brr2 catalyzes an ATP-dependent unwinding of the U4/U6 RNA duplex	?	-	-

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O75643	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography, tag cleavage by TEV protease, follwed by dialysis, another step of nickel affinity chromatography, and ultrafiltration of the flow-through	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + H2O	-	Homo sapiens	ADP + phosphate	-	?
additional information	enzyme Brr2 catalyzes an ATP-dependent unwinding of the U4/U6 RNA duplex	Homo sapiens	?	-	-

Synonyms

Synonyms	Comment	Organism
BRR2	-	Homo sapiens
spliceosomal RNA helicase	-	Homo sapiens

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.0042	-	pH and temperature not specified in the publication	Homo sapiens	3-[5-[(Z)-(5-imino-7-oxo-2-propyl-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid
0.0053	-	pH and temperature not specified in the publication	Homo sapiens	3-[5-[(Z)-(2-cyclohexyl-5-imino-7-oxo-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-6(7H)-ylidene)methyl]furan-2-yl]benzoic acid
0.1	-	above, pH and temperature not specified in the publication	Homo sapiens	(6Z)-6-[[5-(3-chlorophenyl)furan-2-yl]methylidene]-5-imino-2-propyl-5,6-dihydro-7H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-7-one

General Information

General Information	Comment	Organism
evolution	Brr2 is an RNA helicase belonging to the Ski2-like subfamily	Homo sapiens
physiological function	Brr2 is an essential component of spliceosome. Brr2 catalyzes an ATP-dependent unwinding of the U4/U6 RNA duplex, which is a critical step for spliceosomal activation	Homo sapiens