Application | Comment | Organism |
---|---|---|
analysis | CpLDH with APAD+ may be useful as a diagnostic tool for detection of protozoan parasite Cryptosporidium | Cryptosporidium parvum |
Cloned (Comment) | Organism |
---|---|
gene LDH1, recombinant expression of the enzyme in Escherichia coli strain Rosetta (DE3)pLysS, molecular replacement and modelling using the structure of PfLDH, PDB ID 1T2D | Cryptosporidium parvum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme as apo-enzyme and in four ternary complexes, The ternary complexes capture the enzyme bound to NAD/NADH or its 3-acetylpyridine analogue in the cofactor binding pocket, while the substrate binding site is occupied by one of the following ligands: lactate, pyruvate or oxamate. X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution | Cryptosporidium parvum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
gossypol | - |
Cryptosporidium parvum | |
additional information | CpLDH does not display any measurable inhibition to pyruvate concentrations up to at least 20 mM in contrast to many other LDHs | Cryptosporidium parvum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Cryptosporidium parvum | |
0.0018 | - |
(S)-lactate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0085 | - |
APAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0166 | - |
APADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0169 | - |
NADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0303 | - |
NAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.1973 | - |
pyruvate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactate + NAD+ | Cryptosporidium parvum | - |
pyruvate + NADH + H+ | - |
r | |
pyruvate + NADH + H+ | Cryptosporidium parvum | - |
(S)-lactate + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cryptosporidium parvum | Q9GT92 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain Rosetta (DE3)pLysS by ammonium sulfate fractionation, gel filtration, anion exchange chromatography, and ultrafiltration | Cryptosporidium parvum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactate + APAD+ | - |
Cryptosporidium parvum | pyruvate + APADH + H+ | - |
r | |
(S)-lactate + NAD+ | - |
Cryptosporidium parvum | pyruvate + NADH + H+ | - |
r | |
pyruvate + APADH + H+ | - |
Cryptosporidium parvum | (S)-lactate + APAD+ | - |
r | |
pyruvate + NADH + H+ | - |
Cryptosporidium parvum | (S)-lactate + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | the ternary complexes capture the enzyme bound to NAD/NADH or its 3-acetylpyridine analog in the cofactor binding pocket, while the substrate binding site is occupied by one of the following ligands: lactate, pyruvate or oxamate | Cryptosporidium parvum |
Synonyms | Comment | Organism |
---|---|---|
lactate dehydrogenase | - |
Cryptosporidium parvum |
LDH1 | - |
Cryptosporidium parvum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Cryptosporidium parvum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000045 | - |
(S)-lactate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.000055 | - |
NAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.000248 | - |
APAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.00208 | - |
NADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.00209 | - |
pyruvate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.00386 | - |
APADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | 5.5 | reduction of pyruvate | Cryptosporidium parvum |
8.5 | 9.5 | oxidation of lactate | Cryptosporidium parvum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 8 | over 40% of maximal activity for reduction of pyruvate within this range | Cryptosporidium parvum |
8 | 9.5 | 20% of maximal activity for oxidation of lactate at pH 8.0, 90% at pH 8.5, and maximal activity at pH 8.5-9.5, inactive at pH 7.5 | Cryptosporidium parvum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
3-acetyl-pyridine adenine dinucleotide | i.e. APAD+, 3-acetylpyridine adenine dinucleotide, analogue of NAD+ | Cryptosporidium parvum | |
additional information | Cryptosporidium parvum lactate dehydrogenase prefers the acetylpyridine analogue of NADH as a cofactor. NADH/APADH and substrate binding site structures, overview. replacement of the conserved serine residue by Met163 in CpLDH allows a better packing environment for the acetyl of APAD+ | Cryptosporidium parvum | |
NAD+ | - |
Cryptosporidium parvum | |
NADH | - |
Cryptosporidium parvum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0116 | - |
gossypol | pH 5.5, 25°C, recombinant enzyme, with NADH and pyruvate | Cryptosporidium parvum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000033 | - |
(S)-lactate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.00183 | - |
NAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0105 | - |
pyruvate | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.0292 | - |
APAD+ | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.1235 | - |
NADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum | |
0.2326 | - |
APADH | pH 5.5, 25°C, recombinant enzyme | Cryptosporidium parvum |