S-palmitoylation of NOD2 controls its localization to the plasma membrane
Notice bibliographique
Résumé
Nucleotide-binding and oligomerization domain containing protein 2 (NOD2) is a cytosolic pattern-recognition receptor that detects intracellular peptidoglycan (muramyl dipeptide) from bacteria. Membrane association of NOD2 is essential for its ability to activate nuclear factor κB and mitogen-activated protein kinase signaling pathways via the kinase, receptor-interacting serine/threonine-protein kinase 2. The post-translational addition of palmitate to NOD2 results in an acylated protein with increased affinity for membrane bilayers (1Lu Y. Zheng Y. Coyaud É. Zhang C. Selvabaskaran A. Yu Y. Xu Z. Weng X. Chen J.S. Meng Y. Warner N. Cheng X. Liu Y. Yao B. Hu H. et al.Palmitoylation of NOD1 and NOD2 is required for bacterial sensing.Science. 2019; 366: 7Crossref Scopus (60) Google Scholar). Palmitoylation of cysteine residues (shown in orange) at positions C395 and C1033 en face of the structural model (A) based on the crystal structure of rabbit NOD2 (PDB ID: 5IRL) (2Maekawa S. Ohto U. Shibata T. Miyake K. Shimizu T. Crystal structure of NOD2 and its implications in human disease.Nat. Commun. 2016; 7: 11813Crossref PubMed Scopus (102) Google Scholar) is mediated by the protein acyltransferase enzyme zDHHC5 (1Lu Y. Zheng Y. Coyaud É. Zhang C. Selvabaskaran A. Yu Y. Xu Z. Weng X. Chen J.S. Meng Y. Warner N. Cheng X. Liu Y. Yao B. Hu H. et al.Palmitoylation of NOD1 and NOD2 is required for bacterial sensing.Science. 2019; 366: 7Crossref Scopus (60) Google Scholar). The model suggests that this face of the protein is juxtaposed to the plasmalemmal surface. Palmitoylation of the expressed GFP-NOD2WT and GFP-NOD2C395,1033S in HCT116 cells was further characterized using a biorthogonal chemical reporter assay (B) (3Yap M.C. Kostiuk M.A. Martin D.D.O. Perinpanayagam M.A. Hak P.G. Siddam A. Majjigapu J.R. Rajaiah G. Keller B.O. Prescher J.A. Wu P. Bertozzi C.R. Falck J.R. Berthiaume L.G. Rapid and selective detection of fatty acylated proteins using ω-alkynyl-fatty acids and click chemistry.J. Lipid Res. 2010; 51: 1566-1580Abstract Full Text Full Text PDF PubMed Scopus (89) Google Scholar). Here, 15-hexadecynoic acid (15-HDYA), also referred to as alkynyl palmitic acid, was metabolically incorporated into cells. The 15-HDYA covalently attached to the immunocaptured GFP-NOD2 proteins was reacted with azide-PEG3-FLAG via a copper-catalyzed azide–alkyne cycloaddition reaction. The proteins were resolved by SDS-PAGE and subsequently detected by an anti-FLAG antibody. The metabolic label, which was incorporated into NOD2WT and was notably absent in the double cysteine mutant, was removed by treatment with 2.5% hydroxylamine that hydrolyzes the 15-DYA–protein thioester linkage. Finally, the palmitoylation-deficient GFP-NOD2C395,1033S does not localize to the plasma membrane in HCT116 cells (C). Mutations that disrupt S-palmitoylation are associated with severe immunologic and inflammatory diseases such as Crohn’s disease, whereas mutations associated with Blau syndrome demonstrate increased S-palmitoylation and membrane localization (1Lu Y. Zheng Y. Coyaud É. Zhang C. Selvabaskaran A. Yu Y. Xu Z. Weng X. Chen J.S. Meng Y. Warner N. Cheng X. Liu Y. Yao B. Hu H. et al.Palmitoylation of NOD1 and NOD2 is required for bacterial sensing.Science. 2019; 366: 7Crossref Scopus (60) Google Scholar). EQUIPMENT: The following equipment were used: A Zeiss Axiovert 200M microscope with a Hamamatsu ImageEM x2 camera, Yokogawa spinning disc, and a 63× 1.4 NA oil objective. REAGENTS: Alkynyl-palmitate (Click Chemistry Tools), azide-PEG3-FLAG (Jena Bioscience), hydroxylamine, and other copper-catalyzed azide–alkyne cycloaddition reagents (TCEP, CuSO4, and TBTA) were obtained from MilliporeSigma. SOFTWARE: The following software were used: MetaMorph, ImageJ, Python, PyMOL, and AutoDock (4Trott O. Olson A.J. AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading.J. Comput. Chem. 2009; 31: 455-461Google Scholar). This work is supported by the Canadian Institutes of Health Research Project Grants PJT166010 awarded to GDF.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,001 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».