Literature summary for 6.3.3.2 extracted from

Wu, D.; Li, Y.; Song, G.; Cheng, C.; Zhang, R.; Joachimiak, A.; Shaw, N.; Liu, Z.J.
Structural basis for the inhibition of human 5,10-methenyltetrahydrofolate synthetase by N10-substituted folate analogues (2009), Cancer Res., 69, 7294-7301.

Cloned(Commentary)

Cloned (Comment)	Organism
cloning from brain cDNA library, expression of the His-tagged enzyme in Escherichia coli strain BL21	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type MTHFS in a binary complex with ADP, with the N5-iminium phosphate reaction intermediate, or as an enzyme-product complex, hanging drop method at 16°C, 600 nl protein solution, with or without 5 mM 5-formyltetrahydrofolate and 5 mM ATP, are equilibrated against 0.05 ml of reservoir solution containing 100 mM HEPES, pH 6.6, 20 mM MgCl2, 20 mM NiCl2, and 20% w/v PEG 3350, 2 days, X-ray diffraction structure determination and analysis	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant wild-type MTHFS in a binary complex with ADP, with the N5-iminium phosphate reaction intermediate, or as an enzyme-product complex, hanging drop method at 16°C, 600 nl protein solution, with or without 5 mM 5-formyltetrahydrofolate and 5 mM ATP, are equilibrated against 0.05 ml of reservoir solution containing 100 mM HEPES, pH 6.6, 20 mM MgCl2, 20 mM NiCl2, and 20% w/v PEG 3350, 2 days, X-ray diffraction structure determination and analysis

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D154A	site-directed mutagenesis, the mutant shows 0.9% of the wild-type catalytic activity	Homo sapiens
E61A	site-directed mutagenesis, the mutant shows 8.6% of the wild-type catalytic activity	Homo sapiens
F55A	site-directed mutagenesis, the mutant shows 53.6% of the wild-type catalytic activity	Homo sapiens
F85A	site-directed mutagenesis, the mutant shows 72.3% of the wild-type catalytic activity	Homo sapiens
K10A	site-directed mutagenesis, the mutant shows 7.5% of the wild-type catalytic activity	Homo sapiens
K150A	site-directed mutagenesis, the mutant shows wild-type catalytic activity	Homo sapiens
M58A	site-directed mutagenesis, the mutant shows 121.6% of the wild-type catalytic activity	Homo sapiens
M90A	site-directed mutagenesis, the mutant shows 49.2% of the wild-type catalytic activity	Homo sapiens
R145A	site-directed mutagenesis, the mutant shows 2.3% of the wild-type catalytic activity	Homo sapiens
R148A	site-directed mutagenesis, the mutant shows 30.7% of the wild-type catalytic activity	Homo sapiens
R14A	site-directed mutagenesis, the mutant shows 12.9% of the wild-type catalytic activity	Homo sapiens
W109A	site-directed mutagenesis, the mutant shows 39.7% of the wild-type catalytic activity	Homo sapiens
Y152A	site-directed mutagenesis, the mutant shows 24.9% of the wild-type catalytic activity	Homo sapiens
Y153A	site-directed mutagenesis, the mutant shows 3.2% of the wild-type catalytic activity	Homo sapiens
Y83A	site-directed mutagenesis, the mutant shows wild-type catalytic activity	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + 5-formyltetrahydrofolate	Homo sapiens	-	ADP + phosphate + 5,10-methenyltetrahydrofolate	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P49914	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21 by nickel affinity chromatography	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
MCF-7 cell	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + 5-formyltetrahydrofolate	-	Homo sapiens	ADP + phosphate + 5,10-methenyltetrahydrofolate	-	?
ATP + 5-formyltetrahydrofolate	folate binds in a spherical pocket, substrate binding and active site structure, structure-function analysis, detailed overview	Homo sapiens	ADP + phosphate + 5,10-methenyltetrahydrofolate	-	?

Subunits

Subunits	Comment	Organism
More	structure modelling, overview	Homo sapiens

Synonyms

Synonyms	Comment	Organism
5,10-Methenyltetrahydrofolate synthetase	-	Homo sapiens
MTHFS	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens

General Information

General Information	Comment	Organism
malfunction	inhibition of MTHFS in human MCF-7 breast cancer cells arrests the growth of cells	Homo sapiens
physiological function	MTHFS regulates the flow of carbon through the one-carbon metabolic network, which supplies essential components for the growth and proliferation of cells	Homo sapiens