DHHC21 deficiency attenuates renal dysfunction during septic injury

Yang, Xiaoyuan; Zheng, Ethan; Ma, Yonggang; Chatterjee, Victor; Villalba, Nuria; Breslin, Jerome W.; Liu, Ruisheng; Wu, Mack H.; Yuan, Sarah Y.

doi:10.1038/s41598-021-89983-x

Cited by 7 publications

(9 citation statements)

References 56 publications

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“…In recent studies, we identified a significantly increased plasma level of EVs in patients and animals with trauma and sepsis, correlating with exacerbated proinflammatory response and cellular injury ( Beard et al, 2016 ). We also observed greatly increased palmitoylation and DHHC21 activity under inflammatory conditions ( Beard et al, 2016 ; Yang et al, 2021 ). DHHC21, unlike other DHHCs, is localized in plasma membrane, which allows it to dynamically regulate the function of many plasma membrane-associated proteins involved in EV generation, such as G protein-coupled receptors, G proteins, and phospholipases ( Beard et al, 2016 ; Fan et al, 2020 ; Yang et al, 2021 ).…”

Section: Introductionmentioning

confidence: 53%

“…We also observed greatly increased palmitoylation and DHHC21 activity under inflammatory conditions ( Beard et al, 2016 ; Yang et al, 2021 ). DHHC21, unlike other DHHCs, is localized in plasma membrane, which allows it to dynamically regulate the function of many plasma membrane-associated proteins involved in EV generation, such as G protein-coupled receptors, G proteins, and phospholipases ( Beard et al, 2016 ; Fan et al, 2020 ; Yang et al, 2021 ). In the present study, we sought to determine if DHHC21-mediated palmitoylation affects EV production, cargo content, and function during sepsis.…”

Section: Introductionmentioning

confidence: 53%

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Protein palmitoylation regulates extracellular vesicle production and function in sepsis

Yang

Zheng

Chatterjee

et al. 2022

J of Extracellular Bio

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Extracellular vesicles (EVs) are bioactive membrane‐encapsulated particles generated by a series of events involving membrane budding, fission and fusion. Palmitoylation, mediated by DHHC palmitoyl acyltransferases, is a lipidation reaction that increases protein lipophilicity and membrane localization. Here, we report palmitoylation as a novel regulator of EV formation and function during sepsis. Our results showed significantly decreased circulating EVs in mice with DHHC21 functional deficiency (Zdhhc21dep/dep), compared to wild‐type (WT) mice 24 h after septic injury. Furthermore, WT and Zdhhc21dep/dep EVs displayed distinct palmitoyl‐proteomic profiles. Ingenuity pathway analysis indicated that sepsis altered several inflammation related pathways expressed in EVs, among which the most significantly activated was the complement pathway; however, this sepsis‐induced complement enrichment in EVs was greatly blunted in Zdhhc21dep/dep EVs. Functionally, EVs isolated from WT mice with sepsis promoted neutrophil adhesion, transmigration, and neutrophil extracellular trap production; these effects were significantly attenuated by DHHC21 loss‐of‐function. Furthermore, Zdhhc21dep/dep mice displayed reduced neutrophil infiltration in lungs and improved survival after CLP challenges. These findings indicate that blocking palmitoylation via DHHC21 functional deficiency can reduce sepsis‐stimulated production of complement‐enriched EVs and attenuates their effects on neutrophil activity.

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Section: Introductionmentioning

confidence: 53%

Section: Introductionmentioning

confidence: 53%

Protein palmitoylation regulates extracellular vesicle production and function in sepsis

Yang

Zheng

Chatterjee

et al. 2022

J of Extracellular Bio

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“…Our results demonstrate that reducing MLKL and pMLKL acylation via zDHHC21 inactivation restores membrane integrity during necroptosis (Figure 3E). Previous studies showed that zDHHC21 depletion ameliorated septic injury in renal reperfusion 32 and barrier dysfunction in endothelial inflammation 33 , suggesting an important role for zDHHC21 in membrane integrity. Based on these, we decided to focus on the involvement of zDHHC21 in the fate of pMLKL, the functional form of MLKL in necroptosis.…”

Section: Resultsmentioning

confidence: 96%

Acylation of MLKL impacts its function in necroptosis

Pradhan,

Chitkara,

Ramirez

et al. 2023

Preprint

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Mixed lineage kinase domain-like (MLKL) is a key signaling protein of necroptosis. MLKL oligomerizes and translocates to the plasma membrane upon phosphorylation by the necrosome. At the plasma membrane, MLKL induces membrane permeabilization which contributes to the inflammatory activity of necroptosis. Membrane binding of MLKL oligomers is initially initiated by the electrostatic interaction between the protein and membrane phospholipids. We have previously showed that MLKL and its phosphorylated form (pMLKL) are S-acylated during necroptosis and that this acylation increases their membrane binding. In this work, we characterize acylation sites of MLKL and identify multiple cysteines that can undergo acylation with an interesting promiscuity at play. Our results show that MLKL and pMLKL undergo monoacylation, with C184, C269 and C286 being the possible acylation sites. In other words, while there are multiple cysteines that are accessible for acylation, the acylation predominantly occurs at a single cysteine. The molecular interactions that determine the exact acylation site remain unclear. Investigating the S-palmitoyltransferases that might acylate MLKL in necroptosis, we showed that zDHHC21 activity has the strongest effect on pMLKL acylation and reducing the activity of zDHHC21 significantly improved membrane integrity of necroptotic cells. We also observed that inactivation of zDHHC21 reduced the pMLKL levels in the cells, suggesting that blocking the acylation of pMLKL results in reduced membrane binding, destabilization of the protein at the membrane interface and cause its degradation. Finally, through molecular dynamics simulations, we verify the acyl tail of MLKL can insert into the membrane and causes conformational changes to the protein and lipid re-organization of the membrane. At a broader level, our findings shed light onto pMLKL processing and the effect of S-acylation on pMLKL functioning in necroptosis.

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“…Our results demonstrate that reducing MLKL and pMLKL acylation via zDHHC21 inactivation restores membrane integrity during necroptosis (Figure E). Previous studies showed that zDHHC21 depletion ameliorated septic injury in renal reperfusion and barrier dysfunction in endothelial inflammation, suggesting an important role for zDHHC21 in membrane integrity. Based on these, we decided to focus on the involvement of zDHHC21 in the fate of pMLKL, the functional form of MLKL in necroptosis.…”

Section: Resultsmentioning

confidence: 99%

Acylation of MLKL Impacts Its Function in Necroptosis

Pradhan,

Chitkara,

Ramirez

et al. 2024

ACS Chem. Biol.

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Mixed lineage kinase domain-like (MLKL) is a key signaling protein of necroptosis. Upon activation by phosphorylation, MLKL translocates to the plasma membrane and induces membrane permeabilization, which contributes to the necroptosisassociated inflammation. Membrane binding of MLKL is initially initiated by electrostatic interactions between the protein and membrane phospholipids. We previously showed that MLKL and its phosphorylated form (pMLKL) are S-acylated during necroptosis. Here, we characterize the acylation sites of MLKL and identify multiple cysteines that can undergo acylation with an interesting promiscuity at play. Our results show that MLKL and pMLKL undergo acylation at a single cysteine, with C184, C269, and C286 as possible acylation sites. Using all-atom molecular dynamic simulations, we identify differences that the acylation of MLKL causes at the protein and membrane levels. Through investigations of the Spalmitoyltransferases that might acylate pMLKL in necroptosis, we showed that zDHHC21 activity has the strongest effect on pMLKL acylation, inactivation of which profoundly reduced the pMLKL levels in cells and improved membrane integrity. These results suggest that blocking the acylation of pMLKL destabilizes the protein at the membrane interface and causes its degradation, ameliorating the necroptotic activity. At a broader level, our findings shed light on the effect of S-acylation on MLKL functioning in necroptosis and MLKL-membrane interactions mediated by its acylation.

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DHHC21 deficiency attenuates renal dysfunction during septic injury

Cited by 7 publications

References 56 publications

Protein palmitoylation regulates extracellular vesicle production and function in sepsis

Protein palmitoylation regulates extracellular vesicle production and function in sepsis

Acylation of MLKL impacts its function in necroptosis

Acylation of MLKL Impacts Its Function in Necroptosis

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