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Literature summary for 3.5.2.3 extracted from

  • Del Cano-Ochoa, F.; Grande-Garcia, A.; Reverte-Lopez, M.; DAbramo, M.; Ramon-Maiques, S.
    Characterization of the catalytic flexible loop in the dihydroorotase domain of the human multi-enzymatic protein CAD (2018), J. Biol. Chem., 293, 18903-18913 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.2.3

Crystallization (Commentary)

Crystallization (Comment) Organism
several F1563 mutant variants of huDHOase bound to dihydroorotate, X-ray diffraction structure determination and analysis at 1.46-2.12 A resolution Homo sapiens

Protein Variants

Protein Variants Comment Organism
F1563A site-directed mutagenesis, the mutation prevents the closure of the flexible loop and inactivates the enzyme Homo sapiens
F1563L site-directed mutagenesis, the mutation prevents the closure of the flexible loop and inactivates the enzyme Homo sapiens
F1563T site-directed mutagenesis, the mutation prevents the closure of the flexible loop and inactivates the enzyme Homo sapiens
F1563Y site-directed mutagenesis, the mutation prevents enhances the interactions of the flexible loop in the closed position and reduces fluctuations and the reaction rate Homo sapiens
additional information construction of a huDHOase chimera bearing the Escherichia coli DHOase flexible loop, the mutant is inactive Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ dependent on Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60000 80000 gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-carbamoyl-L-aspartate Homo sapiens
-
 (S)-dihydroorotate + H2O
-
 r

Organism

Organism UniProt Comment Textmining
Homo sapiens P27708
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the DHOase reaction is reversible and pH-dependent Homo sapiens ?
-
-
N-carbamoyl-L-aspartate
-
 Homo sapiens (S)-dihydroorotate + H2O
-
 r

Subunits

Subunits Comment Organism
dimer 2 * 30000-40000, SDS-PAGE Homo sapiens
More replacement of the flexible loop weakens the dimerization of huDHOase Homo sapiens

Synonyms

Synonyms Comment Organism
CAD
-
 Homo sapiens
DHOase
-
 Homo sapiens
dihydroorotase domain
-
 Homo sapiens
huDHOase
-
 Homo sapiens

General Information

General Information Comment Organism
evolution despite evolutionary divergence, the CAD DHOase active site components are highly conserved with those in bacterial DHOases, encoded as monofunctional enzymes. An important element for catalysis, conserved from Escherichia coli to humans, is a flexible loop that closes as a lid over the active site. The flexible loop exhibits a two-amino acid signature that is characteristic for each DHOase type Homo sapiens
malfunction a huDHOase chimera bearing the Escherichia coli DHOase flexible loop is inactive, suggesting the presence of distinctive elements in the flexible loop of huDHOase that cannot be replaced by the bacterial sequence. Substitutions of Phe1563 with Ala, Leu, or Thr prevent the closure of the flexible loop and inactivated the protein, whereas substitution with Tyr enhances the interactions of the loop in the closed position and reduced fluctuations and the reaction rate Homo sapiens
metabolism the dihydroorotase (DHOase) domain of the multifunctional protein carbamoyl-phosphate synthetase 2, aspartate transcarbamoylase, and dihydroorotase (CAD) catalyzes the third step in the de novo biosynthesis of pyrimidine nucleotides in animals Homo sapiens
additional information analysis of the catalytic flexible loop in the dihydroorotase domain of the human multi-enzymatic protein CAD, molecular dynamics simulations, overview. Residue Phe1563, a residue absolutely conserved at the tip of the flexible loop in CAD's DHOase domain, is a critical element for the conformational equilibrium between the two catalytic states of the protein. Key role of Phe-1563 in configuring the active site and in promoting substrate strain and catalysis. The flexible loop reaches in toward the active site with N-carbamoyl-L-aspartate bound and is proposed to aid in catalysis by orienting and increasing the electrophilicity of the substrate, excluding water molecules, and stabilizing the transition-state. Then, upon the formation of DHO, the loop moves away from the active site, facilitating product release. As an exception, bacterial type I DHOases present a rigid and shorter loop that interacts minimally with the substrate, requiring the intimate association with ATCase to complete the active site and attain full activity Homo sapiens