Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2
It is known that plant lipid transfer proteins (LTPs) bind a broad spectrum of ligands including fatty acids (FAs), phospho- and glycolipids, acyl-coenzyme A and secondary metabolites. In this work, we used protein−lipid overlay assays to identify new putative LTP ligands. In our experiments, the le...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-11-01
|
| Series: | Membranes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2077-0375/10/11/357 |
| _version_ | 1797547338665820160 |
|---|---|
| author | Daria Melnikova Ivan Bogdanov Tatiana Ovchinnikova Ekaterina Finkina |
| author_facet | Daria Melnikova Ivan Bogdanov Tatiana Ovchinnikova Ekaterina Finkina |
| author_sort | Daria Melnikova |
| collection | DOAJ |
| description | It is known that plant lipid transfer proteins (LTPs) bind a broad spectrum of ligands including fatty acids (FAs), phospho- and glycolipids, acyl-coenzyme A and secondary metabolites. In this work, we used protein−lipid overlay assays to identify new putative LTP ligands. In our experiments, the lentil lipid transfer protein Lc-LTP2 as well as LTPs from other plants were shown to bind phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2). Molecular modeling, 2-p-toluidinonaphthalene-6-sulphonate (TNS) displacement and liposome leakage experiments with Lc-LTP2 and its mutant analogs (R45A, Y80A, R45A/Y80A) were performed to investigate interactions between the protein and PI(4,5)P2. It was shown that PI(4,5)P2 initially interacted with the “bottom” entrance of the protein cavity and after complex formation the large polar head of this ligand was also oriented towards the same entrance. We also found that two highly conserved residues in plant LTPs, Arg45 and Tyr80, played an important role in protein-ligand interactions. Apparently, Arg45 is a key residue for interaction with PI(4,5)P2 during both initial contacting and holding in the protein cavity, while Tyr80 is probably a key amino acid playing an essential role in Lc-LTP2 docking to the membrane. Thus, we assumed that the ability of Lc-LTP2 to bind PI(4,5)P2 suggests the involvement of this protein in plant signal transduction. |
| first_indexed | 2024-03-10T14:42:52Z |
| format | Article |
| id | doaj.art-66e793bdbf5144d9a09465a29b098ef2 |
| institution | Directory Open Access Journal |
| issn | 2077-0375 |
| language | English |
| last_indexed | 2024-03-10T14:42:52Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj.art-66e793bdbf5144d9a09465a29b098ef22023-11-20T21:39:30ZengMDPI AGMembranes2077-03752020-11-01101135710.3390/membranes10110357Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2Daria Melnikova0Ivan Bogdanov1Tatiana Ovchinnikova2Ekaterina Finkina3Science-Educational Center, M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, RussiaScience-Educational Center, M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, RussiaScience-Educational Center, M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, RussiaScience-Educational Center, M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, RussiaIt is known that plant lipid transfer proteins (LTPs) bind a broad spectrum of ligands including fatty acids (FAs), phospho- and glycolipids, acyl-coenzyme A and secondary metabolites. In this work, we used protein−lipid overlay assays to identify new putative LTP ligands. In our experiments, the lentil lipid transfer protein Lc-LTP2 as well as LTPs from other plants were shown to bind phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2). Molecular modeling, 2-p-toluidinonaphthalene-6-sulphonate (TNS) displacement and liposome leakage experiments with Lc-LTP2 and its mutant analogs (R45A, Y80A, R45A/Y80A) were performed to investigate interactions between the protein and PI(4,5)P2. It was shown that PI(4,5)P2 initially interacted with the “bottom” entrance of the protein cavity and after complex formation the large polar head of this ligand was also oriented towards the same entrance. We also found that two highly conserved residues in plant LTPs, Arg45 and Tyr80, played an important role in protein-ligand interactions. Apparently, Arg45 is a key residue for interaction with PI(4,5)P2 during both initial contacting and holding in the protein cavity, while Tyr80 is probably a key amino acid playing an essential role in Lc-LTP2 docking to the membrane. Thus, we assumed that the ability of Lc-LTP2 to bind PI(4,5)P2 suggests the involvement of this protein in plant signal transduction.https://www.mdpi.com/2077-0375/10/11/357lipid transfer proteinlentillipid bindingphosphatidylinositol (4,5)-bisphosphatesignalingmolecular docking |
| spellingShingle | Daria Melnikova Ivan Bogdanov Tatiana Ovchinnikova Ekaterina Finkina Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 Membranes lipid transfer protein lentil lipid binding phosphatidylinositol (4,5)-bisphosphate signaling molecular docking |
| title | Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 |
| title_full | Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 |
| title_fullStr | Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 |
| title_full_unstemmed | Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 |
| title_short | Interaction between the Lentil Lipid Transfer Protein Lc-LTP2 and Its Novel Signal Ligand PI(4,5)P2 |
| title_sort | interaction between the lentil lipid transfer protein lc ltp2 and its novel signal ligand pi 4 5 p2 |
| topic | lipid transfer protein lentil lipid binding phosphatidylinositol (4,5)-bisphosphate signaling molecular docking |
| url | https://www.mdpi.com/2077-0375/10/11/357 |
| work_keys_str_mv | AT dariamelnikova interactionbetweenthelentillipidtransferproteinlcltp2anditsnovelsignalligandpi45p2 AT ivanbogdanov interactionbetweenthelentillipidtransferproteinlcltp2anditsnovelsignalligandpi45p2 AT tatianaovchinnikova interactionbetweenthelentillipidtransferproteinlcltp2anditsnovelsignalligandpi45p2 AT ekaterinafinkina interactionbetweenthelentillipidtransferproteinlcltp2anditsnovelsignalligandpi45p2 |