Cloned (Comment) | Organism |
---|---|
recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Burkholderia multivorans |
Crystallization (Comment) | Organism |
---|---|
purified His6-tagged enzyme in complex with substrate 6-carboxy-5,6,7,8-tetrahydropterin and cofactor S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 2.2-2.6 A resolution | Burkholderia multivorans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | the enzyme uses a bound [4Fe-4S]+ cluster to catalyze the reductive cleavage of S-adenosyl-L-methionine cofactor to generate methionine, a single [4Fe-4S] cluster per monomer. The cluster-binding motifs of each monomer are located at opposite ends of the dimer molecule, of the Fe atoms from the cluster are coordinated by three cysteine residues from the CX14CXPhiC motif. The remaining unique Fe binds the alpha-amino and alpha-carboxyl groups of S-adenosyl-L-methionine, which adopts an anti orientation about the glycosidic bond | Burkholderia multivorans | |
Mg2+ | dependent on, the ligand environment about Mg2+ is pseudo-octahedral, with three water ligands, the substrate C4 carbonyl and C6 carboxylate oxygens, and the hydroxyl of T51 | Burkholderia multivorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
6-carboxy-5,6,7,8-tetrahydropterin | Burkholderia multivorans | - |
7-carboxy-7-carbaguanine + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Burkholderia multivorans | A0A0H3KB22 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Burkholderia multivorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
6-carboxy-5,6,7,8-tetrahydropterin | - |
Burkholderia multivorans | 7-carboxy-7-carbaguanine + NH3 | - |
? | |
6-carboxy-5,6,7,8-tetrahydropterin | the substrate binds perpendicular to the [4Fe-4S]-bound S-adenosyl-L-methionine cofactor, exposing its C6 hydrogen atom for abstraction and generating the binding site for Mg2+, which directly coordinates to the substrate. Substrate binding generates a metal-binding site | Burkholderia multivorans | 7-carboxy-7-carbaguanine + NH3 | - |
? | |
additional information | the enzyme uses a bound [4Fe-4S]+ cluster to catalyze the reductive cleavage of S-adenosyl-L-methionine cofactor to generate methionine and a 5'-deoxyadenosyl radical, which initiates enzymatic transformations requiring H-atom abstraction. To initiate radical chemistry, all AdoMet radical enzymes require the one-electron reduction of the [4Fe-4S] cluster. This electron is typically supplied by a flavodoxin. For the Burkholderia multivorans enzyme, more product is generated using the chemical reductant dithionite | Burkholderia multivorans | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Burkholderia multivorans |
Synonyms | Comment | Organism |
---|---|---|
CDG synthase | - |
Burkholderia multivorans |
queE | - |
Burkholderia multivorans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Burkholderia multivorans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | to initiate radical chemistry, all AdoMet radical enzymes require the one-electron reduction of the [4Fe-4S] cluster. This electron is typically supplied by a flavodoxin. For the Burkholderia multivorans enzyme, more product is generated using the chemical reductant dithionite | Burkholderia multivorans | |
S-adenosyl-L-methionine | the binding site is conserved despite modified enzyme fold | Burkholderia multivorans |
General Information | Comment | Organism |
---|---|---|
evolution | QueE is a member of the radical S-adenosyl-L-methionine superfamily, it varies from the other members in that it contains a hypermodified (beta6/alpha3) protein core and an expanded cluster-binding motif CX14CX2C, structure comparisons, overview | Burkholderia multivorans |
metabolism | the enzyme catalyzes the complex heterocyclic radical-mediated conversion of 6-carboxy-5,6,7,8-tetrahydropterin to 7-carboxy-7-carbaguanine in the third step of the biosynthetic pathway to all 7-deazapurines. 7-Carboxy-7-carbaguanine is the first 7-deazapurine in the biosynthetic pathway and likely the precursor to all naturally occurring 7-deazapurine containing molecules | Burkholderia multivorans |
additional information | substrate binding generates a metal-binding site | Burkholderia multivorans |