Literature summary for 2.8.1.12 extracted from

Daniels, J.N.; Wuebbens, M.M.; Rajagopalan, K.V.; Schindelin, H.
Crystal structure of a molybdopterin synthase-precursor Z complex: insight into its sulfur transfer mechanism and its role in molybdenum cofactor deficiency (2008), Biochemistry, 47, 615-626.

Cloned(Commentary)

Cloned (Comment)	Organism
genes moaE and moaD, expression of His6-tagged MoaE and MoaD in Escherichia coli strain Rosetta (DE3)	Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant apo form molybdopterin synthase and molybdopterin synthase-precursor Z complex using wild-type and mutant K126A MoeE, X-ray diffraction structure determination and analysis, structure modeling with the enzyme from Staphylococcu aureus, overview	Escherichia coli
purified recombinant His-tagged apo form molybdopterin synthase and molybdopterin synthase-precursor Z complex using wild-type and mutant K123A MoeE, 10 mg/ml protein in a buffer containing 50 mM Tris-HCl, pH 8.0, and 50 mM NaCl, mixed with a precipitant consisting of 2.0 M sodium formate and 0.1 M sodium acetate, pH 5.3, at 22°C, 2 days, for the enzyme complex with precursor Z, 18% w/v PEG 8000, 0.1% polyvinylpyrrollidone K15, and 0.1 M Tris-HCl, pH 8.0, at 4°C is used, X-ray diffraction structure determination and analysis at 2.0 A resolution for the apoenzyme and at 2.5 A resolution for the enzyme complex, molecular replacement with MOLREP, modeling using the cyrstal structure of the Escherichia coli MPT synthase, overview	Staphylococcus aureus

Crystallization (Comment)

Organism

purified recombinant apo form molybdopterin synthase and molybdopterin synthase-precursor Z complex using wild-type and mutant K126A MoeE, X-ray diffraction structure determination and analysis, structure modeling with the enzyme from Staphylococcu aureus, overview

Escherichia coli

purified recombinant His-tagged apo form molybdopterin synthase and molybdopterin synthase-precursor Z complex using wild-type and mutant K123A MoeE, 10 mg/ml protein in a buffer containing 50 mM Tris-HCl, pH 8.0, and 50 mM NaCl, mixed with a precipitant consisting of 2.0 M sodium formate and 0.1 M sodium acetate, pH 5.3, at 22°C, 2 days, for the enzyme complex with precursor Z, 18% w/v PEG 8000, 0.1% polyvinylpyrrollidone K15, and 0.1 M Tris-HCl, pH 8.0, at 4°C is used, X-ray diffraction structure determination and analysis at 2.0 A resolution for the apoenzyme and at 2.5 A resolution for the enzyme complex, molecular replacement with MOLREP, modeling using the cyrstal structure of the Escherichia coli MPT synthase, overview

Staphylococcus aureus

Protein Variants

Protein Variants	Comment	Organism
E125K	inactive mutant	Staphylococcus aureus
E128K	inactive mutant	Escherichia coli
K123A	site-directed mutagenesis, mutant of MoaE	Staphylococcus aureus
K126A	site-directed mutagenesis, mutant of MoaE	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	Staphylococcus aureus	-	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	Escherichia coli	-	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P30749	-	-
Staphylococcus aureus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged MoaE and MoaD from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and gel filtration	Staphylococcus aureus

Reaction

Reaction	Comment	Organism	Reaction ID
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O = molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	catalytic mechanism, structure function-relationship, overview	Staphylococcus aureus	a
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O = molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	catalytic mechanism, structure function-relationship, overview	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	-	Staphylococcus aureus	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	-	Escherichia coli	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	via reaction intermediate, overview	Staphylococcus aureus	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?
cyclic pyranopterin phosphate + 2 [molybdopterin-synthase sulfur-carrier protein]-Gly-NH-CH2-C(O)SH + H2O	via reaction intermediate, overview	Escherichia coli	molybdopterin + 2 molybdopterin-synthase sulfur-carrier protein	-	?

Subunits

Subunits	Comment	Organism
heterotetramer	the MPT synthase protein consists of two large (MoaE) and two small (MoaD) subunits with the MoaD subunits located at opposite ends of a central MoaE dimer	Staphylococcus aureus
heterotetramer	the MPT synthase protein consists of two large (MoaE) and two small (MoaD) subunits with the MoaD subunits located at opposite ends of a central MoaE dimer	Escherichia coli

Synonyms

Synonyms	Comment	Organism
MPT synthase	-	Staphylococcus aureus
MPT synthase	-	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Staphylococcus aureus
22	-	assay at room temperature	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Staphylococcus aureus
7.2	-	assay at	Escherichia coli

General Information

General Information	Comment	Organism
metabolism	MPT synthase catalyzes the second step of molybdenum cafactor biosynthesis	Staphylococcus aureus
metabolism	MPT synthase catalyzes the second step of molybdenum cafactor biosynthesis	Escherichia coli
additional information	active site struture and substrate-binding and catalytically important residues, overview	Escherichia coli
additional information	active site struture and substrate-binding and catalytically important residues, overview. Interaction between Glu125 and precursor Z in the enzyme-precursor Z cocrystals	Staphylococcus aureus
physiological function	conversion of the sulfur- and metal-free precursor Z to mylobdopterin by MPT synthase involving the transfer of sulfur atoms from a C-terminal MoaD thiocarboxylate to the C-1' and C-2' positions of precursor Z. In the complex, precursor Z is bound by strictly conserved residues in a pocket at the MoaE dimer interface in close proximity of the C-terminal glycine of MoaD, conformational changes in a loop that participates in interactions with precursor Z	Staphylococcus aureus
physiological function	conversion of the sulfur- and metal-free precursor Z to mylobdopterin by MPT synthase involving the transfer of sulfur atoms from a C-terminal MoaD thiocarboxylate to the C-1' and C-2' positions of precursor Z. In the complex, precursor Z is bound by strictly conserved residues in a pocket at the MoaE dimer interface in close proximity of the C-terminal glycine of MoaD, conformational changes in a loop that participates in interactions with precursor Z	Escherichia coli