Palmitoylation of ACE2 at Cys141, Cys344, and Cys498 facilitates its extracellular vesicles localization

doi:10.16352/j.issn.1001-6325.2025.09.1151

Basic & Clinical Medicine ›› 2025, Vol. 45 ›› Issue (9): 1151-1157.doi: 10.16352/j.issn.1001-6325.2025.09.1151

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Palmitoylation of ACE2 at Cys141, Cys344, and Cys498 facilitates its extracellular vesicles localization

YANG Jingjing¹, MA Yan¹, WANG Yue¹, SUN Yang², LI Pan^2*, GUO Feng^1*

1. Jiangsu Province Key Laboratory of Immunity and Metabolism, Department of Pathogenic Biology and Immunology, Xuzhou Basic Medical School, Xuzhou Medical University, Xuzhou 221004;
2. State Key Laboratory Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical University, Xuzhou 221121, China

Received:2025-06-03 Revised:2025-07-09 Online:2025-09-05 Published:2025-08-27
Contact: ^*PanLi@xzhmu.edu.cn; Feng.Guo@xzhmu.edu.cn

Abstract

Abstract: Objective To identify S-palmitoylation sites on angiotensin-converting enzyme 2(ACE2), the cellular receptor for severe acute respiratory syndrome corona virus(SARS-CoV) and SARS-CoV-2, and to investigate the functional relevance of these modifications in regulating ACE2 localization and activity. Methods An optimized multi-site mutagenesis strategy was performed by simultaneously substitution of all eight cysteine (Cys) residuesin ACE2 by serine to create a non-palmitoylatable mutant (8CS). Then individual cysteine residues were re-introduced one by one. Palmitoylation level of mutants was evaluated using a bioorthogonal click chemistry method to identify palmitoylation-competent residues. Immune-fluorescence staining and extra-cellular vesicle isolation assays were then used to evaluate the impact of specific palmitoylation sites on ACE2 sub-cellular localization. Results An efficient strategy for multi-site palmitoylation site mapping was optimized and successfully identified three critical palmitoylation sites on ACE2: Cys141, Cys344 and Cys498. Functional analyses showed that palmitoylation of these sites significantly promotes the enrichment of ACE2 in extra-cellular vesicles. Conclusions S-palmitoylation at Cys141, Cys344 and Cys498 is essential for the trafficking of ACE2 to extra-cellular vesicles, which suggests a potential regulatory mechanism of its impact on viral receptor presentation and ACE2-associated signaling pathways.

Key words: S-palmitoylation, multi-site mutagenesis, post-translational modification, extra cellular vesicle, angiotensin-converting enzyme 2(ACE2)

CLC Number:

YANG Jingjing, MA Yan, WANG Yue, SUN Yang, LI Pan, GUO Feng. Palmitoylation of ACE2 at Cys141, Cys344, and Cys498 facilitates its extracellular vesicles localization[J]. Basic & Clinical Medicine, 2025, 45(9): 1151-1157.

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Palmitoylation of ACE2 at Cys141, Cys344, and Cys498 facilitates its extracellular vesicles localization

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