Cloned (Comment) | Organism |
---|---|
expression of His-tagged enzyme in Escherichia coli strain Origami (DE3) pLysS | 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | essential divalent cation | Thermotoga maritima |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | Q9X0C3 | - |
- |
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | Q9X0C3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain Origami (DE3) pLysS by nickel affinity chromatography and gel filtration | Thermotoga maritima |
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 | 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 | Comment | Organism |
---|---|---|
guanosine-diphospho-D-mannose pyrophosphorylase | - |
Thermotoga maritima |
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 |