Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58
Lipolysis is an essential metabolic process that releases unesterified fatty acids from neutral lipid stores to maintain energy homeostasis in living organisms. Adipose triglyceride lipase (ATGL) plays a key role in intracellular lipolysis and can be coactivated upon interaction with the protein com...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-01-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227523001645 |
| _version_ | 1797347110220201984 |
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| author | Natalia Kulminskaya Carlos Francisco Rodriguez Gamez Peter Hofer Ines Kathrin Cerk Noopur Dubey Roland Viertlmayr Theo Sagmeister Tea Pavkov-Keller Rudolf Zechner Monika Oberer |
| author_facet | Natalia Kulminskaya Carlos Francisco Rodriguez Gamez Peter Hofer Ines Kathrin Cerk Noopur Dubey Roland Viertlmayr Theo Sagmeister Tea Pavkov-Keller Rudolf Zechner Monika Oberer |
| author_sort | Natalia Kulminskaya |
| collection | DOAJ |
| description | Lipolysis is an essential metabolic process that releases unesterified fatty acids from neutral lipid stores to maintain energy homeostasis in living organisms. Adipose triglyceride lipase (ATGL) plays a key role in intracellular lipolysis and can be coactivated upon interaction with the protein comparative gene identification-58 (CGI-58). The underlying molecular mechanism of ATGL stimulation by CGI-58 is incompletely understood. Based on analysis of evolutionary conservation, we used site directed mutagenesis to study a C-terminally truncated variant and full-length mouse ATGL providing insights in the protein coactivation on a per-residue level. We identified the region from residues N209-N215 in ATGL as essential for coactivation by CGI-58. ATGL variants with amino acids exchanges in this region were still able to hydrolyze triacylglycerol at the basal level and to interact with CGI-58, yet could not be activated by CGI-58. Our studies also demonstrate that full-length mouse ATGL showed higher tolerance to specific single amino acid exchanges in the N209-N215 region upon CGI-58 coactivation compared to C-terminally truncated ATGL variants. The region is either directly involved in protein-protein interaction or essential for conformational changes required in the coactivation process. Three-dimensional models of the ATGL/CGI-58 complex with the artificial intelligence software AlphaFold demonstrated that a large surface area is involved in the protein-protein interaction. Mapping important amino acids for coactivation of both proteins, ATGL and CGI-58, onto the 3D model of the complex locates these essential amino acids at the predicted ATGL/CGI-58 interface thus strongly corroborating the significance of these residues in CGI-58–mediated coactivation of ATGL. |
| first_indexed | 2024-03-08T11:42:40Z |
| format | Article |
| id | doaj.art-aa096cc53e4e4f6a87eb1ee3710210b5 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-03-08T11:42:40Z |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-aa096cc53e4e4f6a87eb1ee3710210b52024-01-25T05:22:33ZengElsevierJournal of Lipid Research0022-22752024-01-01651100491Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58Natalia Kulminskaya0Carlos Francisco Rodriguez Gamez1Peter Hofer2Ines Kathrin Cerk3Noopur Dubey4Roland Viertlmayr5Theo Sagmeister6Tea Pavkov-Keller7Rudolf Zechner8Monika Oberer9Institute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, Austria; BioTechMed Graz, Graz, Austria; BioHealth Field of Excellence, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, Austria; BioTechMed Graz, Graz, Austria; BioHealth Field of Excellence, University of Graz, Graz, AustriaInstitute of Molecular Biosciences, University of Graz, Graz, Austria; BioTechMed Graz, Graz, Austria; BioHealth Field of Excellence, University of Graz, Graz, Austria; For correspondence: Monika ObererLipolysis is an essential metabolic process that releases unesterified fatty acids from neutral lipid stores to maintain energy homeostasis in living organisms. Adipose triglyceride lipase (ATGL) plays a key role in intracellular lipolysis and can be coactivated upon interaction with the protein comparative gene identification-58 (CGI-58). The underlying molecular mechanism of ATGL stimulation by CGI-58 is incompletely understood. Based on analysis of evolutionary conservation, we used site directed mutagenesis to study a C-terminally truncated variant and full-length mouse ATGL providing insights in the protein coactivation on a per-residue level. We identified the region from residues N209-N215 in ATGL as essential for coactivation by CGI-58. ATGL variants with amino acids exchanges in this region were still able to hydrolyze triacylglycerol at the basal level and to interact with CGI-58, yet could not be activated by CGI-58. Our studies also demonstrate that full-length mouse ATGL showed higher tolerance to specific single amino acid exchanges in the N209-N215 region upon CGI-58 coactivation compared to C-terminally truncated ATGL variants. The region is either directly involved in protein-protein interaction or essential for conformational changes required in the coactivation process. Three-dimensional models of the ATGL/CGI-58 complex with the artificial intelligence software AlphaFold demonstrated that a large surface area is involved in the protein-protein interaction. Mapping important amino acids for coactivation of both proteins, ATGL and CGI-58, onto the 3D model of the complex locates these essential amino acids at the predicted ATGL/CGI-58 interface thus strongly corroborating the significance of these residues in CGI-58–mediated coactivation of ATGL.http://www.sciencedirect.com/science/article/pii/S0022227523001645ATGLadipose triglyceride lipasePNPLA2CGI-58comparative gene identification-58ABHD5 |
| spellingShingle | Natalia Kulminskaya Carlos Francisco Rodriguez Gamez Peter Hofer Ines Kathrin Cerk Noopur Dubey Roland Viertlmayr Theo Sagmeister Tea Pavkov-Keller Rudolf Zechner Monika Oberer Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 Journal of Lipid Research ATGL adipose triglyceride lipase PNPLA2 CGI-58 comparative gene identification-58 ABHD5 |
| title | Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 |
| title_full | Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 |
| title_fullStr | Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 |
| title_full_unstemmed | Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 |
| title_short | Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58 |
| title_sort | unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification 58 |
| topic | ATGL adipose triglyceride lipase PNPLA2 CGI-58 comparative gene identification-58 ABHD5 |
| url | http://www.sciencedirect.com/science/article/pii/S0022227523001645 |
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