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Literature summary for 5.6.2.1 extracted from
- Amino acids 39-456 of large subunit and 210-262 of small subunit constitute the minimal, functionally interacting fragments of the unusual heterodimeric topoisomerase IB of Leishmania (2008), Biochem. J., 409, 481 - 489.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| BL21(DE3)pLysS | Leishmania sp. |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D261A LdTOPIS | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
| GST-LdTOPIS delta 1-201 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 1-210 | constitute the minimal interacting fragment. Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 1-210/delta 255-262 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 1-215 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 1-80 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 245-262 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| GST-LdTOPIS delta 255-262 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| K243A LdTOPIS | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
| K436A LdTOPIL | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
| K455A LdTOPIL | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
| LdTOPIL delta 1-39 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| LdTOPIL delta 1-39/delta457-635 | constitute the minimal interacting fragment. Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| LdTOPIL delta 1-99 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| LdTOPIL delta 436-635 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| LdTOPIL delta 457-635 | Generated N-and C-terminal-truncated deletion constructs of of L and S subunit. The heterodimerization between the two fragments is weak and therefore co-purified fragments show reduced DNA binding, cleavage and relaxation properties compared with the wild type enzyme | Leishmania sp. |
| N256A LdTOPIS | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
| N441A LdTOPIL | Site-directed mutagenesis studies identify Lys455 of LdTOPIL and Asp261 of LdTOPIS as two residues involved in subunit interaction. The reconstituted mutant enzymes LdTOPILK436A/S, LdTOPILN441A/S, L/LdTOPISK249A and L/LdTOPISN256A do not show any appreciable change in the relaxation pattern compared with the wild-type enzyme L/S. Reconstitution of L with LdTOPISD261A (SD261A) causes a 15fold decrease in the relaxation activity compared with L/S | Leishmania sp. |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| camptothecin | camptothecin sensitivity fold reduced compared with L/S | Leishmania sp. |  |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| additional information | - |
large subunit (LdTop1L), which contains the non-conserved N-terminal end and a phylogenetically conserved core domain, and of a small subunit (LdTop1S) which harbours the C-terminal region with a characteristic tyrosine residue in the active site | Leishmania sp. |
| 31400 | - |
GST-LdTOPIS 215-262 | Leishmania sp. |
| 31800 | - |
GST-LdTOPIS 210-255 | Leishmania sp. |
| 33000 | - |
GST-LdTOPIS 210-262 | Leishmania sp. |
| 34500 | - |
GST-LdTOPIS 201-262 | Leishmania sp. |
| 46000 | - |
GST-LdTOPIS 80-262 | Leishmania sp. |
| 48700 | - |
6xHis-LdTOPIL 39-456 | Leishmania sp. |
| 51000 | - |
6xHis-LdTOPIL 1-435 | Leishmania sp. |
| 52000 | - |
GST-LdTOPIS 1-255 | Leishmania sp. |
| 53300 | - |
GST-LdTOPIS 1-245 | Leishmania sp. |
| 53500 | - |
6xHis-LdTOPIL1-456 | Leishmania sp. |
| 56000 | - |
GST-LdTOPIS | Leishmania sp. |
| 61700 | - |
6xHis-LdTOPIL 99-635 | Leishmania sp. |
| 68700 | - |
6xHis-LdTOPIL 39-635 | Leishmania sp. |
| 73000 | - |
6xHis-LdTOPIL | Leishmania sp. |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Leishmania sp. | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Proteins are purified through Ni-NTA (Ni2+-nitrilotriacetate)agarose or GST (glutathione transferase)Sepharose 4B column followed by phosphocellulose column (P11 cellulose) | Leishmania sp. |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterodimer | large subunit (LdTOPIL or L) and the catalytic small subunit (LdTOPIS or S) | Leishmania sp. |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Bi-subunit topoisomerase IB | - |
Leishmania sp. |
| DNA topoisomerase I | - |
Leishmania sp. |
| topoisomerase IB | - |
Leishmania sp. |
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