Activating Compound | Comment | Organism | Structure |
---|---|---|---|
PanZ | the Escherichia coli enzyme requires the regulatroy factor PanZ for proteolytic cleavage of the zymogen to form the mature enzyme, the PanZ mutant N45A is unable to complement a panZ deletion strain. Complex formation of the site-directed mutants PanZ(R73A) and PanD(K119A) leads to a complex that still complements the beta-alanine auxotrophy of the DELTApanZ and DELTApanD strains, indicating that catalytically active PanD is formed, but no growth inhibition is observed as a result of PanZ overexpression | Escherichia coli |
Application | Comment | Organism |
---|---|---|
drug development | the PanDZ complex is a target for antibiotic development | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
gene panD, recombinant overexpression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3), subcloning in Escherichia coli strain MG1655. Overexpression-linked growth inhibition is dependent upon CoA-dependent interaction of PanZ with PanD | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
co-crystallization of fully activated PanD and PanZ in complex, in a 10:11 PanD:PanZ ratio (protomer to monomer), hanging drop vapor diffusion method, mixing of 0.003 ml of 9 mg/ml protein in 50 mM Tris, 100 mM NaCl, and 0.1 mM DTT, pH 7.5, with 00.001 ml of reservoir solution containing 200 mM KSCN, 100 mM Bis-Tris propane, pH 6.5, and 20% w/v PEG 3350 at 18°C, X-ray diffraction structure determination and analysis at 1.16 A resolution. The same structure is observed using both room-temperature and cryo-cooled crystals, indicating that the hydrate is formed from the pyruvoyl cofactor and is not an intermediate in the activation reaction. This state is stabilized by a hydrogen bond to the amide of Gly24, which is held in place by interactions with PanZ | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
K115A | site-directed mutagenesis, the mutation is introduced in vitro by overlap extension PCR | Escherichia coli |
K119A | site-directed mutagenesis, the mutation is introduced in vitro by overlap extension PCR. Complex formation of the site-directed mutants PanZ(R73A) and PanD(K119A) leads to a complex that still complements the beta-alanine auxotrophy of the DELTApanZ and DELTApanD strains, indicating that catalytically active PanD is formed, but no growth inhibition is observed as a result of PanZ overexpression | Escherichia coli |
K14A | site-directed mutagenesis, the mutation is introduced in vitro by overlap extension PCR | Escherichia coli |
K53A | site-directed mutagenesis, the mutation is introduced in vitro by overlap extension PCR | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | CoA-dependent interaction of PanZ and PanD. Growth inhibition is due to the CoA-dependent PanD-PanZ interaction and the inhibition occurs at native concentrations of PanD and PanZ in the cell. The production of beta-alanine is feedback-regulated by the PanZ-AcCoA complex | Escherichia coli | |
pentyl pantothenamide | high-potency growth inhibition by pentyl pantothenamide is dependent upon the PanD-PanZ interaction. Substitution of the Escherichia coli panD for the noninteracting Bacillus panD leads to resistance against the inhibitor | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate | Escherichia coli | - |
beta-alanine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
additional information | the Escherichia coli enzyme requires the regulatroy factor PanZ for proteolytic cleavage of the zymogen to form the mature enzyme | Escherichia coli |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain C41(DE3) by sequential immobilized metal affinity chromatography and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate | - |
Escherichia coli | beta-alanine + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ADC | - |
Escherichia coli |
PanD | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyruvoyl cofactor | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
malfunction | both regulatory protein PanZ overexpression-linked beta-alanine auxotrophy and pentyl pantothenamide toxicity are due to formation of the PanDZ complex between enzyme PanD and effector protein PanZ. Formation of such a complex between activated aspartate decarboxylase (PanD) and PanZ leads to sequestration of the pyruvoyl cofactor as a ketone hydrate and demonstrates that both PanZ overexpression-linked beta-alanine auxotrophy and pentyl pantothenamide toxicity are due to formation of this complex. Substitution of the Escherichia coli panD for the noninteracting Bacillus panD suppresses the phenotype | Escherichia coli |
metabolism | enzyme PanD is responsible for the production of beta-alanine in the pantothenate biosynthesis pathway. The production of beta-alanine is feedback-regulated by the PanZ-AcCoA complex | Escherichia coli |
additional information | PanD-PanZ complex three-dimensional structure analysis, overview | Escherichia coli |
physiological function | the PanDZ complex regulates the pantothenate biosynthetic pathway in a cellular context in Escherichia coli by limiting the supply of beta-alanine in response to coenzyme A concentration. Formation of such a complex between activated aspartate decarboxylase (PanD) and regulatory protein PanZ leads to sequestration of the pyruvoyl cofactor as a ketone hydrate. Regulation of PanD is due to CoA-dependent interaction of PanZ and PanD | Escherichia coli |