Literature summary for 6.3.5.2 extracted from

Maruoka, S.; Horita, S.; Lee, W.C.; Nagata, K.; Tanokura, M.
Crystal structure of the ATPPase subunit and its substrate-dependent association with the GATase subunit: a novel regulatory mechanism for a two-subunit-type GMP synthetase from Pyrococcus horikoshii OT3 (2010), J. Mol. Biol., 395, 417-429.

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	all substrates of PH-ATPPase, Mg2+, ATP and XMP, except for ammonia, are required to stabilize the active complex of PH-ATPPase and PH-GATase subunits	Pyrococcus horikoshii

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Pyrococcus horikoshii
genes ph1346 and ph1347, genetic structure of GMPS, overview. Expressionin Escherichia coli strain BL21(DE3)	Pyrococcus horikoshii

Crystallization (Commentary)

Crystallization (Comment)	Organism
ATPPase subunit of the two-subunit-type GMPS, sitting drop vapor diffusion at 5°C, mixing of 0.001 ml of protein solution containing 30 mg/ml protein in Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution containing 30% v/v PEG 400, 100 mM Tris-HCl, pH 8.4, and 200 mM MgCl2, equilibration against 0.1 ml of reservoir solution, 3 weeks, X-ray diffraction structure determination and analysis at 1.8 A resolution	Pyrococcus horikoshii
crystal structure of the ATPPase subunit of the two-subunit-type GMPS, to 1.79 A resolution. ATPPase consists of a N-domain and a C-domain and exists as a homodimer in the crystal and in solution. The N-domain contains an ATP-binding platform called P-loop, whereas the C-domain contains the xanthosine 5'-monophosphate-binding site and also contributes to homodimerization. The glutamine amidotransferase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of PH-ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits	Pyrococcus horikoshii

Crystallization (Comment)

Organism

ATPPase subunit of the two-subunit-type GMPS, sitting drop vapor diffusion at 5°C, mixing of 0.001 ml of protein solution containing 30 mg/ml protein in Tris-HCl, pH 8.0, with 0.001 ml of reservoir solution containing 30% v/v PEG 400, 100 mM Tris-HCl, pH 8.4, and 200 mM MgCl2, equilibration against 0.1 ml of reservoir solution, 3 weeks, X-ray diffraction structure determination and analysis at 1.8 A resolution

Pyrococcus horikoshii

crystal structure of the ATPPase subunit of the two-subunit-type GMPS, to 1.79 A resolution. ATPPase consists of a N-domain and a C-domain and exists as a homodimer in the crystal and in solution. The N-domain contains an ATP-binding platform called P-loop, whereas the C-domain contains the xanthosine 5'-monophosphate-binding site and also contributes to homodimerization. The glutamine amidotransferase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of PH-ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits

Pyrococcus horikoshii

General Stability

General Stability	Organism
all substrates of PH-ATPPase, Mg2+, ATP and XMP, except for ammonia, are required to stabilize the active complex of PH-ATPPase and PH-GATase subunits	Pyrococcus horikoshii

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	as Mg-ATP	Pyrococcus horikoshii

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus horikoshii	O59071	GMP synthase [glutamine-hydrolyzing] subunit A	-
Pyrococcus horikoshii	O59072	strain OT3	-
Pyrococcus horikoshii	O59072	GMP synthase [glutamine-hydrolyzing] subunit B	-
Pyrococcus horikoshii OT-3	O59071	GMP synthase [glutamine-hydrolyzing] subunit A	-
Pyrococcus horikoshii OT-3	O59072	strain OT3	-
Pyrococcus horikoshii OT-3	O59072	GMP synthase [glutamine-hydrolyzing] subunit B	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + xanthosine 5'-phosphate + NH3	overall reaction	Pyrococcus horikoshii	AMP + diphosphate + GMP	-	?
ATP + xanthosine 5'-phosphate + NH3	overall reaction	Pyrococcus horikoshii OT-3	AMP + diphosphate + GMP	-	?
L-glutamine + H2O	GATase half-reaction, structure-activity relationship of the functional subunits, overview	Pyrococcus horikoshii	L-glutamate + NH3	-	?
L-glutamine + H2O	GATase half-reaction, structure-activity relationship of the functional subunits, overview	Pyrococcus horikoshii OT-3	L-glutamate + NH3	-	?
additional information	GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and ATP pyrophosphatase, ATPPase. GATase hydrolyzes glutamine to yield glutamate and ammonia, while ATPPase utilizes ammonia to convert adenyl-XMP into guanosine 5'-monophosphate. The GATase subunit of the two-subunit-type GMPS alone is inactiv	Pyrococcus horikoshii	?	-	?
additional information	GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and ATP pyrophosphatase, ATPPase. GATase hydrolyzes glutamine to yield glutamate and ammonia, while ATPPase utilizes ammonia to convert adenyl-XMP into guanosine 5'-monophosphate. The GATase subunit of the two-subunit-type GMPS alone is inactiv	Pyrococcus horikoshii OT-3	?	-	?
XMP + Mg-ATP2-	ATPPase half-reaction, structure-activity relationship of the functional subunits, overview	Pyrococcus horikoshii	adenyl-XMP + diphosphate + Mg2+	adenyl-XMP is the overall reaction intermediate	?
XMP + Mg-ATP2-	ATPPase half-reaction, structure-activity relationship of the functional subunits, overview	Pyrococcus horikoshii OT-3	adenyl-XMP + diphosphate + Mg2+	adenyl-XMP is the overall reaction intermediate	?

Subunits

Subunits	Comment	Organism
dimer	GMPS consists of two functional units that are present as domains or subunits: glutamine amidotransferase, GATase, and 34 kDa ATP pyrophosphatase, ATPPase. Domain structure and interactions, structure-activity relationship of the functional subunits, overview	Pyrococcus horikoshii
More	the glutamine amidotransferase GATase subunit of the two-subunit-type GMPS alone is inactive, and substrates Mg2+, ATP and XMP of subunit ATP diphosphatase ATPPase except for ammonia are required to stabilize the active complex of ATPPase and GATase subunits	Pyrococcus horikoshii

Synonyms

Synonyms	Comment	Organism
GATase	GMP synthase [glutamine-hydrolyzing] subunit A, glutamine amidotransferase activity	Pyrococcus horikoshii
GMP synthase [glutamine-hydrolyzing] subunit B	ATPPase, ATP diphosphatase activity	Pyrococcus horikoshii
GMPS	-	Pyrococcus horikoshii
Guanosine 5'-monophosphate synthetase	-	Pyrococcus horikoshii
More	GMPS belongs to the class I glutamine amidotransferase, GATase, family	Pyrococcus horikoshii
two-subunit-type GMP synthetase	-	Pyrococcus horikoshii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	assay at	Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Pyrococcus horikoshii

Cofactor

Cofactor	Comment	Organism	Structure
ATP	as Mg-ATP	Pyrococcus horikoshii

General Information

General Information	Comment	Organism
metabolism	GMPS catalyzes the final step of the de novo synthetic pathway of purine nucleotides	Pyrococcus horikoshii