Cloned (Comment) | Organism |
---|---|
expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Paramecium bursaria chlorella virus |
Crystallization (Comment) | Organism |
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purified recombinant His6-tagged wild-type and mutant enzymes, 0.005 ml of 20 mg/ml protein in 10 mM HEPES, pH 7.2, 50 mM NaCl, 1 mM DTT, 0.5 mM EDTA, pH 8.0, and 0.03% v/v Brij-35, with or without 5 mM agmatine, are mixed with an equal volume of the crystallization solution containing 6% PEG 8000, 100 mM imidazole, pH 7.5, 200 mM calcium acetate, sitting drop vapour diffusion, 16°C, crystals generally grow within 24 h, cryoprotection with a solution containing the mother liquor and 25-30% 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis at 1.95 A and 1.8 A resolution for the free and agmatine-bound enzyme, molecular modeling | Paramecium bursaria chlorella virus |
Protein Variants | Comment | Organism |
---|---|---|
T142A | site-directed mutagenesis, structural comparison to the wild-type enzyme, overview | Paramecium bursaria chlorella virus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arginine | Paramecium bursaria chlorella virus | - |
agmatine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paramecium bursaria chlorella virus | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-arginine = agmatine + CO2 | the K148-loop movement may be kinetically linked to the rate-limiting step of product release, active site structure, mechanism | Paramecium bursaria chlorella virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arginine | - |
Paramecium bursaria chlorella virus | agmatine + CO2 | - |
? | |
L-arginine | the key specificity element is the 310-helix that contains and positions substrate-binding residues such as Glu296,mechanism, the 310-helix in Chlorella virus ADC is shifted over 2 A away from the pyridoxal 5'-phosphate cofactor, the K148 loop functions as an active site lid, overview | Paramecium bursaria chlorella virus | agmatine + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the mobile loop, the K148-loop, is observed in a closed, substrate-bound conformation, this loop adopts different conformations throughout the catalytic cycle, overview | Paramecium bursaria chlorella virus |
Synonyms | Comment | Organism |
---|---|---|
ADC | - |
Paramecium bursaria chlorella virus |
More | the enzyme belongs to the group IV of pyridoxal 5'-phosphate-dependent decarboxylases belonging to the alpha/beta barrel structural family | Paramecium bursaria chlorella virus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on, the cofactor forms an interaction with K48 via Schiff base | Paramecium bursaria chlorella virus |