Identification, binding measurements and structure determination of human sirtuin phosphopeptides in complex with human adapter protein 14-3-3σ reveal variations among tight binders and suggest extended binding modes.

Protein glycosylation and phosphorylation are critical post-translational modifications (PTMs) that regulate diverse physiological and pathological pr…

In BriefThe Proteomics Standards Research Group developed a multipathway standard mixture of heavy-labeled phosphopeptides. This mixture contains 131 vetted phosphopeptides that cover biologically int...

AAA ABSTRACT Protein glycosylation and phosphorylation are critical post-translational modifications (PTMs) governing nearly all cellular functions, yet their analysis remains challenging due to reliance on costly and unsustainable enrichment materials. Herein, we report a green synthesis of phosphorylated cellulose nanocrystals (P-CNCs) via one-step phosphoric acid hydrolysis, enabling dual-affinity enrichment of both glycopeptides and phosphopeptides. P-CNCs leverage abundant surface hydroxyl groups for hydrophilic interaction liquid chromatography (HILIC)-based glycopeptide capture, and intrinsic phosphate groups enable direct Ti 4 + chelation (P-CNCs-Ti 4+ ) for phosphopeptide enrichment without chemical derivatization. Using only 1 µL human serum, P-CNCs captured 2,025 N-linked and 2,183 O-GalNAcylated glycopeptides, including 10 previously unreported N-glycosylation chemical modifications. For phosphoproteomics, P-CNCs-Ti 4 + enriched 5,225 phosphopeptides from mouse liver tissue, outperforming commercial TiO 2 and identifying over 100 of 3-phosphoglyceryl modifications in glycolytic enzymes. Comprehensive life cycle assessment demonstrates the environmental sustainability of this approach, achieving a 76% cost reduction compared to commercial materials while significantly lowering the associated environmental footprint. This work pioneers P-CNCs as a sustainable, high-performance platform for multiplexed PTM profiling, bridging glycoproteomics and phosphoproteomics with broad applications in biomarker discovery and metabolic pathway analysis.

Journal of Proteome ResearchDOI: 10.1021/acs.jproteome.4c01091

IonOpticks Aurora Elite 15×75 C18 UHPLC column used in study about Phosphoproteomics optimization by Bortel et al.

Anal. Methods, 2025, Accepted Manuscript DOI: 10.1039/D5AY01446H, PaperJinFeng Zhang, Bo Hu, Huifeng Zhang, Haijiao Zheng, Shufang Yang, Qiong Jia Protein phosphorylation is one of the most critical post-translational modifications for regulating protein activity and function. Direct mass spectrometry (MS) analysis of phosphopeptides presents significant challenges due to their inherently... The content of this RSS Feed (c) The Royal Society of Chemistry