Literature summary for 2.7.7.13 extracted from

Pelissier, M.C.; Lesley, S.A.; Kuhn, P.; Bourne, Y.
Structural insights into the catalytic mechanism of bacterial guanosine-diphospho-D-mannose pyrophosphorylase and its regulation by divalent ions (2010), J. Biol. Chem., 285, 27468-27476.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged enzyme in Escherichia coli strain Origami (DE3) pLysS	Thermotoga maritima

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme in apo form, in complex with substrates mannose-1-phosphate, or with GTP, or bound to the end product GDP-Mannose and Mg2+, sitting drop vapour diffusion method, for apoenzyme: mixing of equal volumes of 20 mg/ml protein in 10 mM Tris, pH 8.0, 150 mM NaCl with reservoir solution containing 35% v/v 2-methyl-2,4-pentanediol, 0.1 M phosphate citrate, pH 7.5, for enzyme complexes: preincubation of 12.5 or 25 mg/ml protein in 10 mM HEPES, pH 7.5, 100 mM NaCl, 10 mM MgCl2, with a 16:1 alpha-D-mannose 1-phosphate or a 8:1 GTP and GDP-Man molar excess of ligand for 30 min at room temperature, protein to well solution ratio of 3:1, the well solution containing 30-35% 2-methyl-2,4-pentanediol, 0.1 M sodium acetate, pH 4.6, 20 mM MgCl2, or in a 2:1 ratio with weel solution containing 30% 2-methyl-2,4-pentanediol, 0.1 M sodium acetate, pH 5.0, 20 mM MgCl2, X-ray diffraction structure determination and analysis at 2.1-2.95 A resolution	Thermotoga maritima

Crystallization (Comment)

Organism

enzyme in apo form, in complex with substrates mannose-1-phosphate, or with GTP, or bound to the end product GDP-Mannose and Mg2+, sitting drop vapour diffusion method, for apoenzyme: mixing of equal volumes of 20 mg/ml protein in 10 mM Tris, pH 8.0, 150 mM NaCl with reservoir solution containing 35% v/v 2-methyl-2,4-pentanediol, 0.1 M phosphate citrate, pH 7.5, for enzyme complexes: preincubation of 12.5 or 25 mg/ml protein in 10 mM HEPES, pH 7.5, 100 mM NaCl, 10 mM MgCl2, with a 16:1 alpha-D-mannose 1-phosphate or a 8:1 GTP and GDP-Man molar excess of ligand for 30 min at room temperature, protein to well solution ratio of 3:1, the well solution containing 30-35% 2-methyl-2,4-pentanediol, 0.1 M sodium acetate, pH 4.6, 20 mM MgCl2, or in a 2:1 ratio with weel solution containing 30% 2-methyl-2,4-pentanediol, 0.1 M sodium acetate, pH 5.0, 20 mM MgCl2, X-ray diffraction structure determination and analysis at 2.1-2.95 A resolution

Thermotoga maritima

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	essential divalent cation	Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
GTP + alpha-D-mannose 1-phosphate	Thermotoga maritima	-	diphosphate + GDP-mannose	-	?
GTP + alpha-D-mannose 1-phosphate	Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589	-	diphosphate + GDP-mannose	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	Q9X0C3	-	-
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589	Q9X0C3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain Origami (DE3) pLysS by nickel affinity chromatography and gel filtration	Thermotoga maritima

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
GTP + alpha-D-mannose 1-phosphate	-	Thermotoga maritima	diphosphate + GDP-mannose	-	?
GTP + alpha-D-mannose 1-phosphate	-	Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589	diphosphate + GDP-mannose	-	?

Subunits

Subunits	Comment	Organism
More	the enzyme has two separate domains: a N-terminal Rossman fold-like domain and a C-terminal left-handed beta-helix domain	Thermotoga maritima

Synonyms

Synonyms	Comment	Organism
guanosine-diphospho-D-mannose pyrophosphorylase	-	Thermotoga maritima

General Information

General Information	Comment	Organism
additional information	the enzyme has two separate domains: a N-terminal Rossman fold-like domain and a C-terminal left-handed beta-helix domain. Two molecules associate into a dimer through a tail-to-tail arrangement of the C-terminal domains. Substrate and product binding are associated with significant changes in the conformation of loop regions lining the active center and in the relative orientation of the two domains, active site structure, overview	Thermotoga maritima
physiological function	the enzyme catalyzes the formation of GDP-Man, a fundamental precursor for protein glycosylation and bacterial cell wall and capsular polysaccharide biosynthesis	Thermotoga maritima