Literature summary for 5.3.1.8 extracted from

Sagurthi, S.R.; Gowda, G.; Savithri, H.S.; Murthy, M.R.
Structures of mannose-6-phosphate isomerase from Salmonella typhimurium bound to metal atoms and substrate: implications for catalytic mechanism (2009), Acta Crystallogr. Sect. D, 65, 724-732.

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of the N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Salmonella enterica subsp. enterica serovar Typhimurium

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme in apoform without metal ion, in the holoform with bound Zn2+, or complexed with bound inhibitor yttrium, or with Zn2+ and fructose 6-phosphate, microbatch method, 0.003 ml of protein solution and crystallization solution are mixed containing 4 mg/ml protein, 0.1 M magnesium acetate, 0.2 M sodium cacodylate, pH 6.5, 20% PEG 8000 and 5% dioxane, addition of 10 mM metal ions and 250 mM fructose 6-phosphate for complexed enzyme, X-ray diffraction structure determination and analysis at 1.7-2.5 A resolution, molecular replacement	Salmonella enterica subsp. enterica serovar Typhimurium

Crystallization (Comment)

Organism

enzyme in apoform without metal ion, in the holoform with bound Zn2+, or complexed with bound inhibitor yttrium, or with Zn2+ and fructose 6-phosphate, microbatch method, 0.003 ml of protein solution and crystallization solution are mixed containing 4 mg/ml protein, 0.1 M magnesium acetate, 0.2 M sodium cacodylate, pH 6.5, 20% PEG 8000 and 5% dioxane, addition of 10 mM metal ions and 250 mM fructose 6-phosphate for complexed enzyme, X-ray diffraction structure determination and analysis at 1.7-2.5 A resolution, molecular replacement

Salmonella enterica subsp. enterica serovar Typhimurium

Inhibitors

Inhibitors	Comment	Organism	Structure
EDTA	complete inhibition at 1 mM, reversible by the addition of divalent metal cations such as Zn2+, Mn2+, Co2+, Mg2+ and Ni2+	Salmonella enterica subsp. enterica serovar Typhimurium
Y3+	-	Salmonella enterica subsp. enterica serovar Typhimurium

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	activates	Salmonella enterica subsp. enterica serovar Typhimurium
Co2+	activates	Salmonella enterica subsp. enterica serovar Typhimurium
Mg2+	best activating divalent metal ion	Salmonella enterica subsp. enterica serovar Typhimurium
Mn2+	activates	Salmonella enterica subsp. enterica serovar Typhimurium
additional information	divalent cations are absolutely required for activity, metal binding increases the thermostability of the enzyme	Salmonella enterica subsp. enterica serovar Typhimurium
Ni2+	activates	Salmonella enterica subsp. enterica serovar Typhimurium
Zn2+	enzyme-bound, Zn2+ binding induces structural order in the loop consisting of residues 50-54. The metal atom appears to play a role in substrate binding and is probably also important for maintaining the architecture of the active site	Salmonella enterica subsp. enterica serovar Typhimurium

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-Mannose 6-phosphate	Salmonella enterica subsp. enterica serovar Typhimurium	-	D-Fructose 6-phosphate	-	r

Organism

Organism	UniProt	Comment	Textmining
Salmonella enterica subsp. enterica serovar Typhimurium	P25081	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Salmonella enterica subsp. enterica serovar Typhimurium

Reaction

Reaction	Comment	Organism	Reaction ID
D-Mannose 6-phosphate = D-fructose 6-phosphate	reaction mechanism, overview. Zn2+ binding induces structural order in the loop consisting of residues 50-54. The metal atom appears to play a role in substrate binding and is probably also important for maintaining the architecture of the active site. Isomerization probably follows a cis-enediol mechanism, overall folding pattern of the central catalytic domain, overview	Salmonella enterica subsp. enterica serovar Typhimurium	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Mannose 6-phosphate	-	Salmonella enterica subsp. enterica serovar Typhimurium	D-Fructose 6-phosphate	-	r

Synonyms

Synonyms	Comment	Organism
MPI	-	Salmonella enterica subsp. enterica serovar Typhimurium

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Salmonella enterica subsp. enterica serovar Typhimurium

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	-	Salmonella enterica subsp. enterica serovar Typhimurium

pH Range

pH Minimum	pH Maximum	Comment	Organism
6.5	9	activity range	Salmonella enterica subsp. enterica serovar Typhimurium