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Literature summary extracted from

  • Huynh, N.; Aye, A.; Li, Y.; Yu, H.; Cao, H.; Tiwari, V.K.; Shin, D.W.; Chen, X.; Fisher, A.J.
    Structural basis for substrate specificity and mechanism of N-acetyl-D-neuraminic acid lyase from Pasteurella multocida (2013), Biochemistry, 52, 8570-8579.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.3.3 recombinant His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pasteurella multocida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.3.3 purified recombinant His-tagged wild-type enzyme, wild-type enzyme with Schiff base and in complex with pyruvate, mutant K164A enzyme, and mutant K164A in complex with N-acetylneuraminic acid and N-glycolylneuraminic acid, hanging drop vapour diffusion method, crystals of wild-type PmNAL from 21% PEG 1000, 150 mM NaCl, and 100 mM Na2HPO4/KH2PO4, pH 6.2. PmNAL K164A mutant in ligand-free from from 30% PEG 200, 100 mM NaCl, and acetate, pH 4.5. Crystals of the PmNAL K164A mutant bound to ligands are grown in 38% PEG 300, 0.01 M CaCl2, and 0.1 M sodium cacodylate, pH 6.5. The sialic acid concentration was 5 mM, X-ray diffraction structure determination and analysis at 1.75-2.10 A resolution, molecular replacement modeling Pasteurella multocida

Protein Variants

EC Number Protein Variants Comment Organism
4.1.3.3 K164A site-directed mutagenesis, determiniation of three structures of the K164A variant: one in the absence of substrates and two binary complexes with N-acetylneuraminic acid and N-glycolylneuraminic acid. Both sialic acids bind to the active site in the open-chain ketone form of the monosaccharide, every hydroxyl group of the linear sugars makes hydrogen bond interactions with the enzyme Pasteurella multocida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.3.3 33700
-
 x * 33700, recombinant His-tagged enzyme, SDS-PAGE Pasteurella multocida

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.3 Pasteurella multocida Q9CKB0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.3.3 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pasteurella multocida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.3 N-acetylneuraminate reversible aldol cleavage of N-acetylneuraminic acid Pasteurella multocida N-acetyl-D-mannosamine + pyruvate
-
 r

Subunits

EC Number Subunits Comment Organism
4.1.3.3 ? x * 33700, recombinant His-tagged enzyme, SDS-PAGE Pasteurella multocida

Synonyms

EC Number Synonyms Comment Organism
4.1.3.3 NAL
-
 Pasteurella multocida