Literature summary for 3.2.1.49 extracted from
Sulzenbacher, G.; Liu, Q.; Bennett, E.; Levery, S.; Bourne, Y.; Ponchel, G.; Clausen, H.; Henrissat, B.
A novel alpha-N-acetylgalactosaminidase family with an NAD+-dependent catalytic mechanism suitable for enzymatic removal of blood group A antigens (2010), Biocatal. Biotransform., 28, 22-32.
No PubMed abstract available
Cloned(Commentary)
Cloned (Comment) |
Organism |
expression in Escherichia coli |
Elizabethkingia meningoseptica |
Crystallization (Commentary)
Crystallization (Comment) |
Organism |
native enzyme and in complex with alpha-N-acetylgalactosamine, to 2.3 and 2.4 A resolution, respectively. Each monomer of NagA consists of two closely associated domains, forming a narrow tunnel in which the NAD+ molecule is anchored. The NAD+ is embedded within a narrow tunnel, almost exclusively shielded from solvent. It is bound with its diphosphate group oriented towards the N-terminus of the first alpha-helix of the N-terminal dinucleotide-binding domain and has both the adenine ring and the nicotinamide ring in the anti conformation. No major structural changes are observed in NagA upon product binding |
Elizabethkingia meningoseptica |
Molecular Weight [Da]
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
48500 |
- |
2 * 48500, SDS-PAGE |
Elizabethkingia meningoseptica |
48500 |
- |
1 * 48500, SDS-PAGE |
Elizabethkingia meningoseptica |
83000 |
- |
gel filtration |
Elizabethkingia meningoseptica |
Organism
Organism |
UniProt |
Comment |
Textmining |
Elizabethkingia meningoseptica |
A4Q8F7 |
member of glycosyl hydrolase family 109 |
- |
Purification (Commentary)
Purification (Comment) |
Organism |
- |
Elizabethkingia meningoseptica |
Reaction
Reaction |
Comment |
Organism |
Reaction ID |
alpha-D-GalNAc-(1->3)-beta-D-GalNAc-(1->3)-alpha-D-Gal-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc-(1<->1)-ceramide + H2O = D-GalNAc + beta-D-GalNAc-(1->3)-alpha-D-Gal-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc-(1<->1)-ceramide |
a hydride at C3, activated by the interaction of the OH-3 hydroxyl with the conserved His228, is abstracted by NAD+ with oxidation of the OH-3 hydroxyl to a ketone. This activates H2 for proton abstraction by Tyr179, accompanied by elimination of the aglycone, thereby generating a 1,2-unsaturated intermediate. Water is then added to the anomeric center, followed by the reduction of the C3 ketone by the on-board NADH, completing the catalytic sequence and restoring NADH to NAD+ and the enzyme to its starting state to bind another substrate |
Elizabethkingia meningoseptica |
|
Substrates and Products (Substrate)
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
2-nitrophenyl-alpha-N-acetylgalactosaminide + H2O |
- |
Elizabethkingia meningoseptica |
2-nitrophenol + alpha-N-acetylgalactosamine |
- |
? |
|
additional information |
enzyme efficiently cleaves blood group A antigen. Unusual catalytic mechanism involving NAD+. NagA might adopt a mechanism similar to that of glycosyl hydrolase family 4 enzymes utilizing an oxidation-elimination-addition-reduction sequence |
Elizabethkingia meningoseptica |
? |
- |
? |
|
Subunits
Subunits |
Comment |
Organism |
dimer |
2 * 48500, SDS-PAGE |
Elizabethkingia meningoseptica |
monomer |
1 * 48500, SDS-PAGE |
Elizabethkingia meningoseptica |
More |
enzyme may exist both in the monomeric and dimeric form, depending on the salt concentration of the buffer solution |
Elizabethkingia meningoseptica |
Synonyms
Synonyms |
Comment |
Organism |
NagA |
- |
Elizabethkingia meningoseptica |
Cofactor
Cofactor |
Comment |
Organism |
Structure |
NAD+ |
binds 1 NAD+ per subunit. The NAD+ cannot dissociate |
Elizabethkingia meningoseptica |
|