Cloned (Comment) | Organism |
---|---|
wild-type and mutants cloned into the pET-28a(+) vector and expressed in Escherichia coli BL21 (DE3) codon plus | Plasmodium falciparum |
Protein Variants | Comment | Organism |
---|---|---|
R187A | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. No inhibition by increasing concentrations of acyl-carrier protein. 3fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R187A/R230A | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. 5fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R187E | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. No inhibition by increasing concentrations of acyl-carrier protein. 4fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R187E/R230E | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. 80fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R187K | no decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R230A | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. No inhibition by increasing concentrations of acyl-carrier protein. 5fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
R230E | kinetic constants (Km and acyl-carrier protein-independent specific activities) remain largely unchanged with respect to wild-type. Exhibits very poor activity in the acyl-carrier protein-dependent spectroscopic assay. No inhibition by increasing concentrations of acyl-carrier protein. 41fold decreased affinity binding to acyl-carrier protein with respect to wild-type | Plasmodium falciparum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acyl-carrier protein | inhibition of wild-type with increasing concentrations | Plasmodium falciparum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.43 | - |
acetoacetyl-CoA | wild-type | Plasmodium falciparum | |
0.44 | - |
acetoacetyl-CoA | mutant R230A | Plasmodium falciparum | |
0.45 | - |
acetoacetyl-CoA | mutant R187A | Plasmodium falciparum | |
0.47 | - |
acetoacetyl-CoA | mutant R187A/R230A | Plasmodium falciparum | |
0.47 | - |
acetoacetyl-CoA | mutant R187E | Plasmodium falciparum | |
0.49 | - |
acetoacetyl-CoA | mutant R230E | Plasmodium falciparum | |
0.58 | - |
acetoacetyl-CoA | mutant R187E/R230E | Plasmodium falciparum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31000 | - |
4 * 31000, SDS-PAGE, wild-type and mutants | Plasmodium falciparum |
110000 | - |
gel filtration, wild-type and mutants | Plasmodium falciparum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutants purified to homogeneity using an Ni2+-nitrilotriacetic acid affinity column | Plasmodium falciparum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2 | - |
mutant R187E/R230E, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
3 | - |
mutant R187A/R230A, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
4 | - |
mutant R230E, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
6.1 | - |
mutant R230A, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
6.3 | - |
mutant R187E, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
6.9 | - |
mutant R187A, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
42.6 | - |
mutant R187E/R230E, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
45.6 | - |
mutant R187A/R230A, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
54.2 | - |
mutant R230E, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
54.5 | - |
mutant R187E, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
55.3 | - |
mutant R230A, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
57.9 | - |
mutant R187A, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
59.8 | - |
wild-type, acyl-carrier protein-independent spectroscopic assay | Plasmodium falciparum |
70.6 | - |
wild-type, acyl-carrier protein-dependent spectroscopic assay | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetoacetyl-CoA + NADPH | wild-type shows less activity with acetoacetyl-CoA than with acetoacetyl-[acyl-carrier protein] | Plasmodium falciparum | ? + NADP+ | - |
? | |
acetoacetyl-[acyl-carrier protein] + NADPH | Arg187 and Arg230 are critical residues for the FabG-acyl-carrier protein interactions, significance of the positively charged/hydrophobic patch located adjacent to the active site cavities of FabG for interactions with acyl carrier protein | Plasmodium falciparum | D-beta-hydroxybutyryl-[acyl-carrier protein] + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 31000, SDS-PAGE, wild-type and mutants | Plasmodium falciparum |
Synonyms | Comment | Organism |
---|---|---|
beta-ketoacyl acyl carrier protein reductase | - |
Plasmodium falciparum |
FabG | - |
Plasmodium falciparum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | 3fold enhancement in acyl-carrier protein binding to wild-type in the presence of NADPH | Plasmodium falciparum |