Biology subjects

Modi, P. K.

Publications and source records attributed to Modi, P. K..

3 recordsLinked to original sources

bioRxiv2026

Temporal phosphoproteomics reveals rapid restoration of kinase signaling by Glycyrrhiza glabra in a rotenone-induced Parkinson disease model

Parkinsons disease is a progressive neurodegenerative disorder associated with mitochondrial dysfunction, oxidative stress, impaired autophagy, and dysregulated cellular signaling pathways. Although Glycyrrhiza glabra has been reported to exhibit neuroprotective properties, the early phosphorylation-mediated signaling mechanisms underlying its protective effects remain poorly understood. In this study, we employed a Tandem Mass Tag (TMT)-based temporal quantitative phosphoproteomic approach to investigate early signaling events associated with Glycyrrhiza glabra-mediated neuroprotection in a rotenone-induced in vitro PD model. Differentiated IMR-32 neuronal cells were treated with rotenone alone or in combination with Glycyrrhiza glabra extract, and phosphoproteomic alterations were analyzed at 2, 5, 15, and 30 minutes using liquid chromatography coupled with tandem mass spectrometer. Temporal phosphoproteomic analysis identified 6,424 phosphopeptides corresponding to 2,368 phosphoproteins and 5,468 phosphorylation sites. Comparative analysis revealed extensive phosphorylation rewiring induced by rotenone and restoration of several dysregulated phosphorylation events following Glycyrrhiza glabra co-treatment. More than 130 phosphoproteins and multiple kinase-associated signaling pathways were dynamically regulated across the temporal conditions. Kinase enrichment analysis identified restoration of several critical kinases, including AKT1, MTOR, MAPK1/3, PRKACA, PRKCD, and GSK3A/B, which are associated with neuronal survival, stress adaptation, and autophagy. Integrated pathway and kinase-substrate interaction analyses further revealed enrichment of AMPK signaling, FOXO signaling, receptor tyrosine kinase signaling, RNA processing, and cell-cycle regulatory pathways. Notably, several spliceosome-associated phosphoproteins demonstrated dynamic phosphorylation changes during the early neuroprotective response. Collectively, this study provides a detailed temporal phosphoproteomic landscape of early signaling events associated with Glycyrrhiza glabra-mediated neuroprotection and highlights kinase-driven signaling pathways that may represent potential therapeutic targets in Parkinsons disease.

neuroscience↗

bioRxiv2026

Phosphoproteomic profiling reveals signaling pathways modulated by Annona muricata leaf extract in oral adenosquamous carcinoma cells

Phosphorylation driven dysregulation of intracellular signaling networks is a central feature of cancer initiation, progression, and therapeutic resistance. Although Annona muricata leaf extracts have demonstrated anticancer activity across multiple experimental models, the underlying molecular mechanisms particularly at the level of phosphorylation dependent signaling remain poorly understood. In this study, we employed a tandem mass tag TMT-based quantitative phosphoproteomic approach to systematically characterize signaling alterations induced by methanolic Annona muricataleaf extract (AME) in oral squamous cell carcinoma (OSCC) CAL-27 cells. Functional assays revealed that AME significantly inhibited cell proliferation, migration, and clonogenic survival. Global phosphoproteomic profiling identified 6,362 phosphopeptides corresponding to 1,964 unique phosphorylation sites across nearly 7,000 phosphoproteins. AME treatment induced widespread, time-dependent hypophosphorylation, indicating a coordinated suppression of oncogenic signaling networks. Pathway and network analyses revealed marked inhibition of signaling pathways associated with key oncogenic kinases, including cyclin-dependent kinases (CDKs), mitogen-activated protein kinases (MAPKs), and signaling modules linked to EGFR and mTOR pathways. Kinase-substrate enrichment and kinome mapping further demonstrated reduced inferred activity of CDK and MAPK driven signaling, accompanied by suppression of cell cycle progression, mitosis, and checkpoint regulation. Collectively, these findings demonstrate that AME induces systems-level remodeling of phosphorylation dependent signaling networks, enforcing a growth-restrictive cellular state in OSCC cells. This study highlights quantitative phosphoproteomics as a powerful strategy for dissecting natural product mediated regulation of oncogenic signaling and provides mechanistic insight into the anticancer potential of Annona muricata.

cancer biology↗

bioRxiv2026

Serial enrichment based quantitative proteomics enables deep and multiplexed profiling of post-translational modifications

Post-translational modifications (PTMs) play central roles in regulating protein function, localization, stability, and signaling networks. Comprehensive characterization of multiple PTMs, however, remains technically challenging due to limited sample availability, enrichment, incompatibilities, and analytical complexity. Here, we present an integrated serial enrichment workflow that enables quantitative profiling of multiple PTMs from a single biological sample. Using a streamlined strategy that combines StageTip-based fractionation with sequential immunoaffinity and metal oxide affinity enrichment, we achieved robust identification and quantification of lysine acetylation, lysine succinylation, phosphotyrosine, and global phosphorylation within the same sample. Application of this workflow resulted in deep proteome coverage alongside high-confidence PTM site identification with minimal sample loss and high reproducibility. Importantly, serial enrichment preserved PTM specificity and enabled comparative quantitative analysis across modification types. This approach provides a practical and scalable solution for integrated multi-PTM analysis, facilitating comprehensive interrogation of proteome regulation under limited sample conditions and offering broad applicability to systems biology, disease profiling, and translational proteomics studies.

systems biology↗