Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap) DOI Creative Commons
Michael T. Forrester, Jacob R. Egol, Aleksandra Tata

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 27, 2024

Proteins undergo reversible S -acylation via a thioester linkage in vivo. -palmitoylation, modification by C16:0 fatty acid, is common that mediates critical protein-membrane and protein-protein interactions. The most widely used assays, including acyl-biotin exchange acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage the subsequent labeling nascent thiol. These assays generally require >500 micrograms protein input material per sample numerous reagent removal washing steps, making them laborious ill-suited for high throughput low applications. To overcome these limitations, we devised Acyl-Trap, suspension trap-based assay utilizes thiol-reactive quartz to enable buffer hydroxylamine-mediated -acyl enrichment. We show method compatible with protein-level detection -acylated proteins (e.g. H-Ras) as well site identification quantification using on-trap isobaric LC-MS/MS from little 20 input. In mouse brain, Acyl-Trap identified 279 reported sites 1298 previously unreported putative sites. Also described are conditions long-term hydroxylamine storage, which streamlines assay. More generally, serves proof-of-concept PTM-tailored traps suitable both traditional chemoproteomic workflows.

Language: Английский

Cryo-EM structure of SARS-CoV-2 postfusion spike in membrane DOI Open Access
Wei Shi, Yongfei Cai, Haisun Zhu

et al.

Nature, Journal Year: 2023, Volume and Issue: 619(7969), P. 403 - 409

Published: June 7, 2023

Language: Английский

Citations

59
Protein lipidation in health and disease: molecular basis, physiological function and pathological implication DOI Creative Commons
Yuan Yuan, Peiyuan Li, Jianghui Li

et al.

Signal Transduction and Targeted Therapy, Journal Year: 2024, Volume and Issue: 9(1)

Published: March 15, 2024

Abstract Posttranslational modifications increase the complexity and functional diversity of proteins in response to complex external stimuli internal changes. Among these, protein lipidations which refer lipid attachment are prominent, primarily encompassing five types including S-palmitoylation, N-myristoylation, S-prenylation, glycosylphosphatidylinositol (GPI) anchor cholesterylation. Lipid plays an essential role regulation trafficking, localisation, stability, conformation, interactions signal transduction by enhancing hydrophobicity. Accumulating evidence from genetic, structural, biomedical studies has consistently shown that lipidation is pivotal broad physiological functions inextricably linked a variety diseases. Decades dedicated research have driven development wide range drugs targeting lipidation, several agents been developed tested preclinical clinical studies, some which, such as asciminib lonafarnib FDA-approved for therapeutic use, indicating represents promising strategy. Here, we comprehensively review known regulatory enzymes catalytic mechanisms various types, outline impact on physiology disease, highlight potential targets progress, aiming provide comprehensive reference future research.

Language: Английский

Citations

27
Protein post-translational modification in SARS-CoV-2 and host interaction DOI Creative Commons

Nana Cheng,

Mingzhu Liu,

Wanting Li

et al.

Frontiers in Immunology, Journal Year: 2023, Volume and Issue: 13

Published: Jan. 13, 2023

SARS-CoV-2 can cause lung diseases, such as pneumonia and acute respiratory distress syndrome, multi-system dysfunction. Post-translational modifications (PTMs) related to are conservative pathogenic, the common PTMs glycosylation, phosphorylation, acylation. The glycosylation of mainly occurs on spike (S) protein, which mediates entry virus into cells through interaction with angiotensin-converting enzyme 2. utilizes glycans cover its epitopes evade immune response S protein. Phosphorylation nucleocapsid (N) protein improves selective binding viral RNA promotes replication transcription, thereby increasing load in host. Succinylated N membrane(M) proteins synergistically affect particle assembly. regulates affinity for other genome acetylation. acetylated envelope (E) interacts bromodomain-containing 2/4 influence host response. Both palmitoylation myristoylation sites infectivity. Papain-like protease is a domain NSP3 that dysregulates inflammation by deubiquitination impinges IFN-I antiviral responses deISGylation. Ubiquitination ORF7a inhibits IFN-α signaling blocking STAT2 phosphorylation. methylation inhibit formation stress granules promote RNA, promoting production particles. macrodomain reverse ADP-ribosylation proteins, cascade IFN core, intracellular SARS-CoV-2. On whole, have fundamental roles entry, replication, assembly, Mutations various variants, lead changes at corresponding sites, different biological effects. In this paper, we reviewed effects cells, whose application inform strategies inhibiting infection facilitating treatment vaccine development COVID-19.

Language: Английский

Citations

28
Insight into the role of clathrin‐mediated endocytosis inhibitors in SARS‐CoV‐2 infection DOI
Samar Sami Alkafaas,

Abanoub Mosaad Abdallah,

Soumya Ghosh

et al.

Reviews in Medical Virology, Journal Year: 2022, Volume and Issue: 33(1)

Published: Nov. 7, 2022

Abstract Emergence of SARS‐CoV‐2 variants warrants sustainable efforts to upgrade both the diagnostic and therapeutic protocols. Understanding details cellular molecular basis virus–host cell interaction is essential for developing variant‐independent options. The internalization SARS‐CoV‐2, into lung epithelial cells, mediated by endocytosis, especially clathrin‐mediated endocytosis (CME). Although vaccination gold standard strategy against viral infection, selective inhibition endocytic proteins, complexes, associated adaptor proteins may present a strategy. clathrin and/or dynamins are most important involved in CME, other mechanisms dynamin independent rely on proteins. Moreover, implicates some subcellular structures, like plasma membrane, actin lysosomes. Also, physiological conditions, such as pH ion concentrations, represent an additional factor that mediates these events. Accordingly, related potential targets small molecules inhibit endocytosis‐mediated entry. This review summarizes using molecules, targeting key participating clathrin‐dependent ‐independent antiviral drugs infection. takes two approaches. first outlines role inhibitors preventing entry its mechanism action, whereas second computational analysis was implemented investigate selectivity common endocytosis. revealed remdesivir, methyl‐β‐cyclodextrin, rottlerin, Bis‐T can effectively clathrin, HMG‐CoA reductase, actin, I GTPase more potent inhibiting than chloroquine. CME infection remain understudied.

Language: Английский

Citations

30
Glycosylation and S-palmitoylation regulate SARS-CoV-2 spike protein intracellular trafficking DOI Creative Commons

Chih-Feng Tien,

Wan-Ting Tsai,

Chun-Hwa Chen

et al.

iScience, Journal Year: 2022, Volume and Issue: 25(8), P. 104709 - 104709

Published: July 4, 2022

Post-translational modifications (PTMs), such as glycosylation and palmitoylation, are critical to protein folding, stability, intracellular trafficking, function. Understanding regulation of PTMs SARS-CoV-2 spike (S) could help the therapeutic drug design. Herein, VSV vector was used produce S pseudoviruses examine roles 611LYQD614 cysteine-rich motifs in maturation virus infectivity. Our results show that 611LY612 mutation alters trafficking reduces cell surface expression level. It also changes pattern decreases pseudovirus The contains four clusters with I II main palmitoylation sites. Mutations disrupt from ER-to-Golgi, suppress production, reduce spike-mediated membrane fusion activity. Taken together, orchestrate processing for protein-mediated infection.

Language: Английский

Citations

26
CellPalmSeq: A curated RNAseq database of palmitoylating and de-palmitoylating enzyme expression in human cell types and laboratory cell lines DOI Creative Commons
Angela R. Wild, Peter W. Hogg, Stéphane Flibotte

et al.

Frontiers in Physiology, Journal Year: 2023, Volume and Issue: 14

Published: Jan. 24, 2023

The reversible lipid modification protein S-palmitoylation can dynamically modify the localization, diffusion, function, conformation and physical interactions of substrate proteins. Dysregulated is associated with a multitude human diseases including brain metabolic disorders, viral infection cancer. However, diverse expression patterns genes that regulate palmitoylation in broad range cell types are currently unexplored, their commonly used lines workhorse basic preclinical research often overlooked when studying dependent processes. We therefore created CellPalmSeq (https://cellpalmseq.med.ubc.ca), curated RNAseq database interactive webtool for visualization across single types, bulk tissue, cancer laboratory non-human lines. This resource will allow exploration these patterns, revealing important insights into cellular physiology disease, aid line selection interpretation results processes depend on S-palmitoylation.

Language: Английский

Citations

15
Modulators for palmitoylation of proteins and small molecules DOI

Zeshuai Fan,

Yuchen Hao,

Yidan Huo

et al.

European Journal of Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 271, P. 116408 - 116408

Published: April 12, 2024

Language: Английский

Citations

5
Palmitoylation of SARS-CoV-2 Envelope protein is central to virus particle formation DOI
Z. W. Wang,

Manman Qiu,

Yue Ji

et al.

Journal of Virology, Journal Year: 2024, Volume and Issue: 98(10)

Published: Sept. 17, 2024

ABSTRACT The Envelope (E) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an integral structural in the virus particles. However, its role assembly virions and underlying molecular mechanisms are yet to be elucidated, including whether function E regulated by post-translational modifications. In present study, we report that SARS-CoV-2 palmitoylated at C40, C43, C44 palmitoyltransferases zDHHC3, 6, 12, 15, 20. Mutating these three cysteines serines (C40/43/44S) reduced stability protein, decreased interaction with proteins Spike, Membrane, Nucleocapsid, thereby inhibited production virus-like particles (VLPs) VLP-mediated luciferase transcriptional delivery. Specifically, C40/43/44S mutation density VLPs. Collectively, results demonstrate palmitoylation vital for IMPORTANCE this systematically examined biochemistry demonstrated required particle (VLP) maintaining normal density. These suggest central proper morphogenesis VLPs densities viral infectivity. This study presents a significant advancement understanding how assembling supports palmitoyl acyltransferases can potential therapeutic targets development inhibitors.

Language: Английский

Citations

4
The fusion peptide of the spike protein S2 domain may be a mimetic analog of β-coronaviruses and serve as a novel virus-host membrane fusion inhibitor DOI
Abass Alao Safiriyu,

Afaq Hussain,

Nikesh Dewangan

et al.

Antiviral Research, Journal Year: 2025, Volume and Issue: unknown, P. 106144 - 106144

Published: March 1, 2025

Language: Английский

Citations

0
Palmitoylated COX-2Cys555 reprogramed mitochondrial metabolism in pyroptotic inflammatory injury in patients with post-acute COVID-19 syndrome DOI Creative Commons

Jia‐Shen Wu,

Chi-Yu Xu,

Su-Min Mo

et al.

Journal of Advanced Research, Journal Year: 2025, Volume and Issue: unknown

Published: May 1, 2025

Language: Английский

Citations

0