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Literature summary for 2.5.1.56 extracted from

  • Joseph, D.D.; Jiao, W.; Kessans, S.A.; Parker, E.J.
    Substrate-mediated control of the conformation of an ancillary domain delivers a competent catalytic site for N-acetylneuraminic acid synthase (2014), Proteins, 82, 2054-2066.
    View publication on PubMed
Search for more literature for 2.5.1.56

Crystallization (Commentary)

Crystallization (Comment) Organism
by small angle X-ray scattering and structure analysis, PDB ID 1XUZ Neisseria meningitidis

Protein Variants

Protein Variants Comment Organism
E134A site-directed mutagenesis Neisseria meningitidis
E282A site-directed mutagenesis Neisseria meningitidis
E282A/T285A site-directed mutagenesis Neisseria meningitidis
additional information generation of a truncated variant of enzyme NANAS which lacks the C-terminal AFPL domain, G272Term by inserting a stop codon. The truncation is made at residue Gly272 prior to the linker region Neisseria meningitidis
T285A site-directed mutagenesis Neisseria meningitidis
T285F site-directed mutagenesis Neisseria meningitidis

General Stability

General Stability Organism
phosphoenolpyruvate has a major effect on the thermal stability of the wild-type enzyme requiring the C-terminal AFPL domain Neisseria meningitidis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic analysis of wild-ype and mutant enzymes, no kinetic activity can be observed for mutant G272Term with phosphoenolpyruvate, binding constants for wild-type, overview Neisseria meningitidis

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ required, increases the enzyme stability, binding of Mn2+ is associated with a change in the average conformation of enzyme NmeNANAS Neisseria meningitidis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O Neisseria meningitidis
-
 phosphate + N-acetylneuraminate
-
 ?
phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O Neisseria meningitidis MC58
-
 phosphate + N-acetylneuraminate
-
 ?

Organism

Organism UniProt Comment Textmining
Neisseria meningitidis H2VFG5 serotype B, gene synC
-
Neisseria meningitidis MC58 H2VFG5 serotype B, gene synC
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information binding of phosphoenolpyruvate is associated with a change in the average conformation of enzyme NmeNANAS causing changes in flexibility in both AFPL domain and ManNAc binding loop Neisseria meningitidis ?
-
 ?
additional information binding of phosphoenolpyruvate is associated with a change in the average conformation of enzyme NmeNANAS causing changes in flexibility in both AFPL domain and ManNAc binding loop Neisseria meningitidis MC58 ?
-
 ?
phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O
-
 Neisseria meningitidis phosphate + N-acetylneuraminate
-
 ?
phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O
-
 Neisseria meningitidis MC58 phosphate + N-acetylneuraminate
-
 ?

Subunits

Subunits Comment Organism
homodimer the enzyme comprises two distinct domains, an N-terminal catalytic (beta/alpha)8 barrel linked to a C-terminal antifreeze protein-like (AFPL) domain. Loss of the AFPL domain destabilizes the dimeric form of the enzyme nd renders it inactive. The AFPL domain plays a critical role for both the catalytic function and quaternary structure stability of N-acetylneuraminic acid synthase. The enzyme mutant G272Term lacking the AFPL domain is partly monomeric Neisseria meningitidis

Synonyms

Synonyms Comment Organism
NANAS
-
 Neisseria meningitidis
NmeNANAS
-
 Neisseria meningitidis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 the wild-type apoenzyme has a higher melting temperature of 42.6°C compared to the mutant G272Term apoenzyme with 36.5°C. Unlike the wild-type enzyme, the melting temperature of mutant G272Term in the presence of Mn2+ remains consistent and is not influenced by the addition of different ligand combinations. The melting temperature of wild-type NmeNANAS increases slightly in the presence of Mn2+ Neisseria meningitidis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Neisseria meningitidis

General Information

General Information Comment Organism
malfunction a truncated variant of the enzyme lacking the C-terminal AFPL domain is soluble and catalytically inactive, loss of the AFPL domain destabilizes the dimeric form of the enzyme. The AFPL domain plays a critical role for both the catalytic function and quaternary structure stability of N-acetylneuraminic acid synthase. phosphoenolpyruvate has a major effect on the thermal stability of the enzyme and loss of the AFPL domain in mutant G272Term eliminates this property Neisseria meningitidis
additional information a complex hydrogen-bonding relay links the roles of the catalytic and AFPL domains across subunit boundaries, analyzed by small angle X-ray scattering, molecular dynamics simulations, and amino acid substitutions, overview Neisseria meningitidis