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High-resolution crystal structures of Streptococcus pneumoniae nicotinamidase with trapped intermediates provide insights into the catalytic mechanism and inhibition by aldehydes

French, J.B.; Cen, Y.; Sauve, A.A.; Ealick, S.E.; Biochemistry 49, 8803-8812 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.1.19
expressed in Escherichia coli Rosetta 2(DE3) pLysS cells
Streptococcus pneumoniae
Crystallization (Commentary)
EC Number
Crystallization
Organism
3.5.1.19
a trapped nicotinoyl-thioester complexed with wild type enzyme and C136S mutant in complex with nicotinamide, hanging drop vapor diffusion method, using 18-22% (w/v) polyethylene glycol 3350, 0.2-0.3 M NaCl and 0.2 M sodium malonate, pH 6.3, at 18°C
Streptococcus pneumoniae
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
C136S
inactive
Streptococcus pneumoniae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
5-methoxynicotinaldehyde
competitive inhibitor
Streptococcus pneumoniae
3.5.1.19
nicotinaldehyde
competitive inhibitor
Streptococcus pneumoniae
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
Co2+
addition of Co2+ after metal removement results in 13% activity
Streptococcus pneumoniae
3.5.1.19
Fe2+
addition of Fe2+ after metal removement results in 13% activity
Streptococcus pneumoniae
3.5.1.19
Mn2+
addition of Mn2+ after metal removement results in 27% activity
Streptococcus pneumoniae
3.5.1.19
additional information
addition of Fe3+ or Ni2+ after metal removement results in no detectable activity
Streptococcus pneumoniae
3.5.1.19
Zn2+
contains zinc, highest level of activity in the presence of Zn2+
Streptococcus pneumoniae
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.1.19
21400
-
4 * 21400, calculated from amino acid sequence
Streptococcus pneumoniae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.1.19
nicotinamide + H2O
Streptococcus pneumoniae
-
nicotinate + NH3
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.1.19
Streptococcus pneumoniae
Q97PM2
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.1.19
Ni-NTA resin column chromatography
Streptococcus pneumoniae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.1.19
nicotinamide + H2O
-
718883
Streptococcus pneumoniae
nicotinate + NH3
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.5.1.19
homotetramer
4 * 21400, calculated from amino acid sequence
Streptococcus pneumoniae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
expressed in Escherichia coli Rosetta 2(DE3) pLysS cells
Streptococcus pneumoniae
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
3.5.1.19
a trapped nicotinoyl-thioester complexed with wild type enzyme and C136S mutant in complex with nicotinamide, hanging drop vapor diffusion method, using 18-22% (w/v) polyethylene glycol 3350, 0.2-0.3 M NaCl and 0.2 M sodium malonate, pH 6.3, at 18°C
Streptococcus pneumoniae
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
C136S
inactive
Streptococcus pneumoniae
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
5-methoxynicotinaldehyde
competitive inhibitor
Streptococcus pneumoniae
3.5.1.19
nicotinaldehyde
competitive inhibitor
Streptococcus pneumoniae
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
Co2+
addition of Co2+ after metal removement results in 13% activity
Streptococcus pneumoniae
3.5.1.19
Fe2+
addition of Fe2+ after metal removement results in 13% activity
Streptococcus pneumoniae
3.5.1.19
Mn2+
addition of Mn2+ after metal removement results in 27% activity
Streptococcus pneumoniae
3.5.1.19
additional information
addition of Fe3+ or Ni2+ after metal removement results in no detectable activity
Streptococcus pneumoniae
3.5.1.19
Zn2+
contains zinc, highest level of activity in the presence of Zn2+
Streptococcus pneumoniae
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.1.19
21400
-
4 * 21400, calculated from amino acid sequence
Streptococcus pneumoniae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.1.19
nicotinamide + H2O
Streptococcus pneumoniae
-
nicotinate + NH3
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
Ni-NTA resin column chromatography
Streptococcus pneumoniae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.1.19
nicotinamide + H2O
-
718883
Streptococcus pneumoniae
nicotinate + NH3
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.1.19
homotetramer
4 * 21400, calculated from amino acid sequence
Streptococcus pneumoniae