Identification of substrates of ZDHHC19 in mouse haploid spermatids using click chemistry
Ontology highlight
ABSTRACT:
INSTRUMENT(S):
ORGANISM(S): Mus Musculus (mouse)
TISSUE(S): Testis
SUBMITTER: 
Jiaoyang Li
LAB HEAD: Ruibao Ren
PROVIDER: PXD057727 | Pride | 2025-09-10
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| RRB-CC-KO-NEG.raw | Raw | |||
| RRB-CC-KO-P.raw | Raw | |||
| RRB-CC-WT-NEG.raw | Raw | |||
| RRB-CC-WT-P.raw | Raw | |||
| checksum.txt | Txt |
Items per page: 5 1 - 5 of 6 |
Similar Datasets
Project description:Reversible palmitoylation is catalyzed by palmitoyltransferase(PAT) from the zinc-finger DHHC domain-containing protein (ZDHHC) family. The Zdhhc19 knockout male mice showed infertility and Zdhhc19 deficient sperm showed severe morphological defects. To identify the candidate substrates of ZDHHC19 in haploid spermatids, we carried out label free mass spectrometry (MS) combined with resin-assisted capture (RAC) methods. This study reveals the role of palmitoylation mediated by ZDHHC19 during spermiogenesis.
2025-09-10 | PXD057735 | Pride
Project description:Although sex differences in the brain are prevalent, the knowledge about mechanisms underlying sex-related effects on normal and pathological brain functioning is rather poor. It is known that female and male brains differ in size and connectivity. Moreover, those differences are related to neuronal morphology and activity, synapse molecular organization, synaptic plasticity, and molecular signalling pathways. Among different processes assuring proper synapse function are posttranslational modifications, and among them, S-palmitoylation emerges as a crucial mechanism underlying synaptic integrity. Protein S-palmitoylation (S-PALM) refers to the covalent attachment of palmitic acid (C16:0) to cysteine residue(s) via the formation of a thioester bond. Protein S-PALM is governed by a family of PATs, also known as DHHC proteins. Here we focused on the sex-related functional importance of DHHC7 acyltransferase. Under physiological conditions, DHHC7 emerges of particular interest because it palmitoylates various synaptic proteins involved in the regulation of cellular polarity and proliferation as well as in stress and anxiety-related pathology. Moreover, DHHC7 is responsible for S-PALM of sex steroid receptors. Functionally, S-PALM of these receptors regulates their trafficking to the plasma membrane as well as their rapid responses to sex steroid hormones. Using mass spectrometry-based method PANIMoni we identified sex-dependent differences in the S-palmitoylation of synaptic proteins, potentially involved in the regulation of passive membrane properties and excitability (e.g. potassium voltage-gated channels), synaptic transmission (e.g. subunits of glutamate or kainate receptors, voltage-dependent calcium channels) as well as signalling proteins involved in structural plasticity of dendritic spines (e.g. G-proteins and Rab GTPase). Furthermore, to determine a mechanistic source for sex-dependent changes in protein S-palmitoylation we used synaptoneurosomes from DHHC-7 knock-out -/- (DHHC7 KO) female and male mice. Our data showed sex-dependent action of DHHC-7 acyltransferase. Such discovery will facilitate the development of novel and targeted treatments congruous with biological sex.
2021-06-23 | PXD025286 | Pride
Project description:Protein S-palmitoylation is a reversible post-translational modification which adding 16-carbon palmitate to specific cysteines and contributes to variable biological functions such as protein trafficking, scaffolding and signaling. ZDHHC13 is one of the members of DHHC-containing palmitoyl-acyltransferase (PAT) family of enzyme. The Zdhhc13 deficient mice show the most severe phenotype with amyloidosis, alopecia, and osteoporosis. Here we used quantitative proteomic approach, combining resin-assisted capture (RAC) method and mass spectrometry-based label free strategy to identify the candidate Zdhhc13 substrates from mouse liver tissue.
2017-09-29 | PXD003039 | Pride
Project description:ZDHHC S-acyltransferases catalyze the transfer of fatty acyl groups, primarily palmitate, to cysteine residues of substrate proteins in all eukaryotes. The 23 human ZDHHCs are known to S-acylate an extensive network of proteins essential to normal physiology, including many which are dysregulated in disease. Current technologies for whole proteome analyses of S-acylation do not directly associate a specific ZDHHC with its substrates, making it difficult to link a ZDHHC to specific mechanisms or disease phenotypes. Here, we present a solution to this challenge through development and validation of the first ZDHHC chemical genetic systems. Intact cells expressing a ZDHHC engineered to carry a ‘hole’ mutation in the lipid binding domain were metabolically labeled with a synthetic, chemically-tagged fatty acid probe carrying a matching ‘bump’, resulting in selective fatty acid probe transfer to substrates of the mutant ZDHHC. ZDHHC/probe pairs were exemplified for efficient and selective transfer to ZDHHCs 3, 7, 11, 15 and 20, and adapted to chemical proteomic analysis for ZDHHCs 7, 15 and 20, generating the first de novo substrate profiles for these ZDHHC isozymes across three different cell lines. These data reveal extensive ZDHHC-specific substrate sets across more than 300 total identified substrates, in addition to substrates common between cell lines and between ZDHHCs, and several functionally diverse ZDHHC20 substrates were validated in targeted assays and the sites of S-acylation determined. The chemical genetic platform described here offers a novel and versatile approach to explore ZDHHC substrates and S-acylation biology, with the potential to encompass ZDHHCs across a wide range of eukaryotic organisms. This submission contains data relating to the determination of changes in S-acylated proteins using YnPal and DHHC20 CRISPR Cas9 KO cells, alongside determining the interactome of ZDHHC20 using TurboID.
2023-10-08 | PXD032378 | Pride
Project description:Attachment of lipids to proteins is a key form of protein modification which intersects with all areas of cellular physiology 1 . Of these, the most pervasive form of protein lipidation is protein S-acylation, also commonly known as protein palmitoylation. Protein palmitoylation is a post-translational modification whereby long chain fatty acids, most typically the 16-carbon palmitic acid, is attached to a cytosol-facing cysteine through a thioester linkage. More than 10% of the proteome is targeted by this modification2, which is catalyzed by members of the zDHHC family of integral membrane enzymes. In humans, there are 23 members of the zDHHC family localized at a variety of organellar membranes as well as the plasma membrane that catalyze protein palmitoylation. Despite having been discovered more than twenty years ago 3 4 the chemistry and biology of many of the zDHHC members remain poorly understood and for several members, few substrates have been characterized. Intriguingly, there are no primary sequence determinants that dictate sites of protein palmitoylation. Cysteines that are proximal to the membrane have a high propensity of being palmitoylated.
2023-10-04 | PXD041525 | Pride
Project description:The Post-translational modification of Hippo/YAP pathway members plays an important role in regulating the activation of Hippo/YAP pathway and its effects in tumors. S-Palmitoylation, also known as S-acylation,is the most common protein Lipid modification, mainly composed of palmitoyl transferases (PATs) from the DHHC (Asp His His Cys) family Catalysis. The results showed that ZDHHC15 mediated palmitoylation of KIBRA up-regulated the expression of YAP downstream target genes, and regulated the invasion function of breast cancer and ovarian cancer cells. The research results will reveal new regulatory mechanisms of the Hippo pathway
2024-12-17 | GSE235254 | GEO
Project description:The Post-translational modification of Hippo/YAP pathway members plays an important role in regulating the activation of Hippo/YAP pathway and its effects in tumors. S-Palmitoylation, also known as S-acylation, is the most common protein Lipid modification, mainly composed of palmitoyl transferases (PATs) from the DHHC (Asp His His Cys) family Catalysis. The results showed that ZDHHC15 mediated palmitoylation of KIBRA up-regulated the expression of YAP downstream target genes, and regulated the invasion function of breast cancer and ovarian cancer cells. The research results will reveal new regulatory mechanisms of the Hippo pathway
2024-12-17 | GSE235258 | GEO
Project description:Protein S-palmitoylation, a dynamic and reversible post-translational modification involving the attachment of palmitate to cysteine residues, is a key regulator of protein functionality and cellular signaling. Dysregulation of this modification has emerged as a critical driver of cancer progression, influencing oncogenes and signaling pathways. Among the 23 DHHC palmitoyl transferases responsible for catalyzing S-palmitoylation, aberrant expression of specific members is linked to tumorigenesis and development, underscoring their potential as promising therapeutic targets. However, the cancer-specific roles and substrates of individual DHHC enzymes remain poorly characterized. In this study, we identified DHHC9 as a crucial regulator of adenocarcinoma progression, including colorectal and lung cancers. Functional studies demonstrated that DHHC9 knockdown profoundly inhibited cell migration in vitro and tumor metastasis in vivo. Proteomic analysis revealed STRN4, a core component of the STRIPAK complex, as a DHHC9-specific substrate whose palmitoylation promotes YAP nuclear localization and activates Hippo pathway effectors CCN1, CCN2, and ANKRD1, driving cancer cell migration. Notably, we discovered two small molecules, treprostinil and 10-HCPT, as potent DHHC9 inhibitors that effectively suppressed adenocarcinoma cell migration. Our findings illuminate a pivotal role for DHHC9 in adenocarcinoma metastasis and unveil its potential as a highly promising therapeutic target in cancer treatment, opening new avenues for intervention in palmitoylation-driven malignancies.
2025-09-15 | PXD060045 | Pride
Project description:Myelin sheath is an important structure to maintain normal functions of the nerves in central nervous system. Protein palmitoylation has been established as a sorting determinant for the transport of myelin-forming proteins to the myelin membrane. However, its function in the regulation of oligodendrocyte development remains unknown. Here, we show that an Asp-His-His-Cys (DHHC) motif-containing palmitoyl acyltransferases, DHHC5, is involved in the control of oligodendrocyte development. Loss of Zdhhc5 in oligodendrocytes inhibits myelination and remyelination by a reduction on total myelinating oligodendrocyte population. STAT3 is the primary substrate for DHHC5 palmitoylation in oligodendrocytes. Zdhhc5 ablation reduces STAT3 palmitoylation, and then suppresses STAT3 phosphorylation and activation. As a result of the decreased STAT3 transactivity, the transcription of the myelin-related and anti-apoptosis genes is therefore inhibited, leading to suppressed oligodendrocyte development and myelination. Our findings demonstrate the key role DHHC5 in the control of myelinogenesis.
2024-02-04 | GSE169247 | GEO
Project description:Myelin sheath is an important structure to maintain normal functions of the nerves in central nervous system. Protein palmitoylation has been established as a sorting determinant for the transport of myelin-forming proteins to the myelin membrane. However, its function in the regulation of oligodendrocyte development remains unknown. Here, we show that an Asp-His-His-Cys (DHHC) motif-containing palmitoyl acyltransferases, DHHC5, is involved in the control of oligodendrocyte development. Loss of Zdhhc5 in oligodendrocytes inhibits myelination and remyelination by a reduction on total myelinating oligodendrocyte population. STAT3 is the primary substrate for DHHC5 palmitoylation in oligodendrocytes. Zdhhc5 ablation reduces STAT3 palmitoylation, and then suppresses STAT3 phosphorylation and activation. As a result of the decreased STAT3 transactivity, the transcription of the myelin-related and anti-apoptosis genes is therefore inhibited, leading to suppressed oligodendrocyte development and myelination. Our findings demonstrate the key role DHHC5 in the control of myelinogenesis.
2024-02-04 | GSE169248 | GEO