Cloned (Comment) | Organism |
---|---|
recombinant expression of truncated enzyme mutants from plasmid pMALcHT, encoding a maltose-binding protein (MBP) followed by a linker region composed of a tobacco etch virus (TEV) protease cut site and a six histidine affinity tag, in Escherichia coli strain BL21-Star (DE3), recombinnat expression of wild-type and truncated mutant enzymes in lipoylation-deficient Escherichia coli strain JEG3. Structure comparisons, overview | Plasmodium falciparum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant lipoyl-PfLipL1D243-279 complex, mixing of 0.001 ml of 5 mg/ml protein, 1.2 mol equiv of lipoate, and excess ATP with 0.001 ml of reservoir solution containing 100 mM HEPES, pH 7.0, 1.5 M (NH4)2SO4, and 20% ethylene glycol, 20°C, method optimization, X-ray diffraction structure determination and analysis at 2.32 A resolution | Plasmodium falciparum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of truncated mutants of enzyme PfLipL1, PfLipL1DELTA259-269, PfLipL1DELTA254-274, and PfLipL1D249-279 | Plasmodium falciparum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Plasmodium falciparum | 5739 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Plasmodium falciparum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-lipoate + a [lipoyl-carrier protein]-L-lysine | Plasmodium falciparum | - |
a [lipoyl-carrier protein]-N6-(lipoyl)lysine + AMP + diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | Q8IEG9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged truncated mutant enzymes from Escherichia coli strain BL21-Star (DE3) by nickel affinity chromatography, MBP tag cleavage by TEV protease, cation exchange chromatography, and gel filtration | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-lipoate + a [lipoyl-carrier protein]-L-lysine | - |
Plasmodium falciparum | a [lipoyl-carrier protein]-N6-(lipoyl)lysine + AMP + diphosphate | - |
? | |
additional information | cell-based lipoylation assays using wild-type and truncated mutant enzymes recombinantly expressed in lipoylation-deficient Escherichia coli strain JEG3 | Plasmodium falciparum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the lipoate-bound PfLipL1DELTA243-279 structure consists of a large N-terminal domain (NTD, residues 21-276), a linker region lacking defined secondary structure (residues 277-289), and a small C-terminal domain (CTD, residues 290-370). The NTD contains two beta-sheets, a large mixed beta-sheet made up of seven beta-strands (beta1, beta2, beta6, beta7, beta8, beta9, and beta10) and a small mixed-sheet made up of three strands (beta3, beta4, and beta5). There are six alpha-helical elements surrounding the beta-sheets (alpha16). The CTD consists of one anti-parallel beta-sheet made up of three strands (beta11, beta12, and beta13) and three alpha-helices (alpha7, alpha8, and alpha9). The two monomers in the asymmetric unit of similar conformations | Plasmodium falciparum |
Synonyms | Comment | Organism |
---|---|---|
LipL1 | - |
Plasmodium falciparum |
lipoate-bound lipoate ligase 1 | - |
Plasmodium falciparum |
PfLipL1 | - |
Plasmodium falciparum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Plasmodium falciparum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Plasmodium falciparum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Plasmodium falciparum |
General Information | Comment | Organism |
---|---|---|
evolution | lipoate protein ligase 1 (PfLipL1) is an ATP-dependent ligase that belongs to the biotin/lipoate A/B protein ligase family (PFAM PF03099) | Plasmodium falciparum |
physiological function | PfLipL1 is the only known canonical lipoate ligase in Plasmodium falciparum and functions as a redox switch between two lipoylation routes in the parasite mitochondrion | Plasmodium falciparum |