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Literature summary for 2.7.7.85 extracted from

  • Heidemann, J.L.; Neumann, P.; Dickmanns, A.; Ficner, R.
    Crystal structures of the c-di-AMP-synthesizing enzyme CdaA (2019), J. Biol. Chem., 294, 10463-10470 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.85

Application

Application Comment Organism
drug development the single enzyme is an attractive target for the development of antibiotic compounds Listeria monocytogenes EGD

Cloned(Commentary)

Cloned (Comment) Organism
gene dacA, recombinant expression of N-terminally GST-tagged truncated enzymes DELTA300cdaA and DELTA300cdaA-Y187A, which lacks the transmembrane domain, in Escherichia coli strain BL21(DE3) Listeria monocytogenes EGD

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant truncated wild-type and mutant enzymes CdaA in the apo state, in the post-catalytic state with bound c-di-AMP and catalytic Co2+ ions, as well as in a complex with AMP, X-ray diffraction structure determination and analysis at 2.0-2.8 A resolution Listeria monocytogenes EGD

Protein Variants

Protein Variants Comment Organism
additional information generation of the DELTA300cdaA allele, which lacks the transmembrane domain Listeria monocytogenes EGD
Y187A site-directed mutagenesis, the mutation to Ala leads to drastic loss of enzymatic activity Listeria monocytogenes EGD

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates slightly, can only partly substitute for Mn2+. Co2+ is coordinated by the phosphate moiety and the carboxylate group of Glu224 as well as the carboxylate group of Asp171 and the imidazole ring nitrogen of His170 of the symmetry-related subunit Listeria monocytogenes EGD
Mn2+ activates, required for catalysis Listeria monocytogenes EGD

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 ATP Listeria monocytogenes EGD
-
 2 diphosphate + cyclic di-3',5'-adenylate
-
 ?
2 ATP Listeria monocytogenes EGD ATCC BAA-679
-
 2 diphosphate + cyclic di-3',5'-adenylate
-
 ?

Organism

Organism UniProt Comment Textmining
Listeria monocytogenes EGD Q8Y5E4
-
-
Listeria monocytogenes EGD ATCC BAA-679 Q8Y5E4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged truncated enzymes DELTA300cdaA and DELTA300cdaA-Y187A from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography, dialysis, tag cleavage, and another step of glutathione chromatography Listeria monocytogenes EGD

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ATP
-
 Listeria monocytogenes EGD 2 diphosphate + cyclic di-3',5'-adenylate
-
 ?
2 ATP
-
 Listeria monocytogenes EGD ATCC BAA-679 2 diphosphate + cyclic di-3',5'-adenylate
-
 ?

Subunits

Subunits Comment Organism
homodimer the CdaA-CdaA dimer interface buries about 605 A2 of the accessible surface area (7.3%) and is stabilized by six hydrogen bonds and two salt bridges. Additional interactions between the monomers are mediated by the ligand bound to the active site Listeria monocytogenes EGD

Synonyms

Synonyms Comment Organism
c-di-AMP-synthesizing enzyme
-
 Listeria monocytogenes EGD
CdaA
-
 Listeria monocytogenes EGD

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Listeria monocytogenes EGD

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Listeria monocytogenes EGD

General Information

General Information Comment Organism
evolution cyclic di-AMP (c-di-AMP) is the only second messenger known to be essential for bacterial growth. It is mainly found in Gram-positive bacteria, including pathogenic bacteria like Listeria monocytogenes. CdaA is the sole diadenylate cyclase in Listeria monocytogenes Listeria monocytogenes EGD
malfunction the essential role of Tyr187 is confirmed by mutation to Ala, leading to drastic loss of enzymatic activity Listeria monocytogenes EGD
additional information flexibility of tyrosine 187 side chain involved in locking the adenine ring after ATP binding, residue Y187 is essential for enzyme activity. Structure analysis of the active site of dimeric CdaA with bound c-di-AMP, overview. In the monomeric CdaA-AMP complex, the tyrosine is rotated inward at the active site and stacks on the adenine in an almost coplanar orientation. In contrast, in the dimeric c-di-AMP complex, the tyrosine side chain is flipped outward, as the Thr202 side chain of the other subunit packs against the adenine ring Listeria monocytogenes EGD