Dataset Information

A metabolic complex of acyltransferase enzymes involved in tomato acylsugar biosynthesis

ABSTRACT: Specialized metabolites mediate diverse plant-environment interactions. Recent work has begun to enzymatically characterize entire plant specialized metabolic pathways; however, little is known about how different pathway components organize and interact within the cell. Here we use acylsugars - a class of specialized metabolites – to explore metabolic complex formation. In Solanum lycopersicum (tomato) four trichome-localized acylsugar acyltransferases (SlASAT1-4) sequentially add acyl chains to a sucrose core leading to accumulation of tri and tetraacylated sucroses. Confocal microscopy demonstrates that tomato ASATs localize to distinct subcellular compartments, including the mitochondria, cytosol, and endoplasmic reticulum. To explore pairwise protein-protein interactions in acylsugar biosynthesis, we used various techniques relying on different interaction principles, including co-immunoprecipitation, split luciferase assays, and bimolecular fluorescence complementation all demonstrating pairwise ASAT interactions. Following transient expression of SlASAT1-4 in N. benthamiana, we were able to pull down a complex consisting of SlASAT1-4, which was confirmed through proteomics. Size exclusion chromatography of the SlASAT pulldown suggests a heteromultimeric complex of around 300 kDa. This study sheds light on the metabolic coordination of acylsugar biosynthesis through formation of a metabolic complex coordinating production of chemical defenses.

INSTRUMENT(S):

ORGANISM(S): Nicotiana Tabacum (common Tobacco) Solanum Lycopersicum

SUBMITTER: Craig Schenck

LAB HEAD: Craig Schenck

PROVIDER: PXD072183 | Pride | 2026-04-27

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20241209_153548_Varun_Craig_Dec2024.sne	Other
	Varun_1_S2-C1_1_6281.d.zip	Other
	Varun_2_S2-C2_1_6282.d.zip	Other
	Varun_3_S2-C3_1_6283.d.zip	Other
	Varun_4_S2-C4_1_6284.d.zip	Other

Items per page:

1 - 5 of 8

Similar Datasets

Project description:In this study, we employed a hypothesis-free and data-driven analytical approach to investigate the CSF proteome in 29 patients with SLE. Herein, our aim was to explore the CSF proteome with data-independent acquisition mass spectrometry (DIA-MS) and cluster SLE patients based on their CSF proteomic patterns. In addition, we aimed to investigate the relationship between protein patterns and a comprehensive clinical dataset including demographic variables, medical history, clinical rheumatologic and neurologic disease manifestations, neurocognitive functionality, cerebral MRI and laboratory measurements.The proteomic data was used for sample clustering and clusters were analyzed for clinical dataset variance. Proteins were clustered in modules using Weighted Gene Co-expression Correlation Network Analysis (WGCNA) and modules were biologically characterized and analyzed for correlation to the clinical dataset. Three patient clusters were identified. Cluster 1 was characterized by the highest frequency of nephritis, depression, and cognitive dysfunction. Cluster 2 showed the highest frequency of alopecia and SSA-antibodies, and a low frequency of cognitive impairment. Cluster 3 had a higher frequency of autonomic neuropathy and lupus headache. Six protein modules were identified (M1-M6). The modules were characterized by nervous tissue proteins (M1), CNS lipoproteins (M2), macrophage proteins (M3), plasma proteins (M4), immunoglobulins (M5), and intracellular metabolic proteins (M6). Module 1 and M2 proteins were most abundant in patient cluster 1 and correlated with nephritis, depression and cognitive impairment. Increased abundance of M4 and M5 proteins were most distinct in patient cluster 2 and inversely correlated to cognitive impairment and brain atrophy. We conclude that patients clustered by their CSF proteomic pattern had different disease phenotypes. Nephritis and neuronal damage defined the group with higher levels of neuronal proteins in CSF, which may suggest shared pathogenetic pathways in SLE affecting the kidney and CNS.

Project description:Background: Extracellular matrix remodeling mechanisms are understudied in cardiac development and congenital heart defects. Two similar matrix-degrading metalloproteases, ADAMTS1 and ADAMTS5, are extensively co-expressed during mouse cardiac development. The mouse mutants of each have mild cardiac anomalies, but their combined genetic inactivation is precluded by tight linkage. We coupled Adamts1 inactivation with pharmacologic ADAMTS5 blockade to uncover stage-specific cooperative roles and mechanisms in mouse cardiac development. Methods: ADAMTS5 blockade was achieved in Adamts1 null mouse embryos using an activity-blocking monoclonal antibody during distinct developmental windows covering myocardial compaction or cardiac septation and outflow tract rotation. Synchrotron imaging, RNA in situ hybridization, immunofluorescence microscopy and electron microscopy were used to determine the impact on cardiac development and compared to Gpc6 and ADAMTS-cleavage resistant mouse mutants. Mass spectrometry-based N-terminomics was used to identify relevant substrates. Results: Combined inactivation of ADAMTS1 and ADAMTS5 prior to 12.5 days of gestation led to dramatic accumulation of versican-rich cardiac jelly and inhibited formation of compact and trabecular myocardium, which we also observed in mice with ADAMTS cleavage-resistant versican. Subsequently, combined knockout impaired outflow tract development and ventricular septal closure, generating a tetralogy of Fallot-like defect independently of versican proteolysis. N-terminomics of combined ADAMTS knockout and wild-type hearts identified a cleaved glypican-6 peptide only in the wild-type and showed that ADAMTS1 and ADAMTS5 each cleaved glypican-6. Paradoxically, ADAMTS1 and ADAMTS5 inactivated hearts lacked glypican-6 despite unaltered Gpc6 transcription. Gpc6-/- mice demonstrated similar rotational defects as the combined ADAMTS knockout and both had reduced Hedgehog signaling. Conclusions: ADAMTS1 and ADAMTS5 ensure proper cardiac development via cleavage of distinct proteoglycans, each with independent roles in cardiac development. Whereas versican clearance in cardiac jelly is required for proper ventricular cardiomyogenesis, glypican-6 cleavage may activate/stabilize this cell-surface proteoglycan which is required for Hedgehog signaling during outflow tract development.

Dataset Information

A metabolic complex of acyltransferase enzymes involved in tomato acylsugar biosynthesis

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets