Dataset Information

Deep quantitative proteomics of North American Pacific coast star tunicate

ABSTRACT: The first proteomics analysis of a colonial tunicate (Botryllus schlosseri) was carried out to support development of a DIA assay library and quantitative proteomics for this species. Deep proteome coverage (up to 4930 protein groups and 20,984 unique peptides in a single sample) was achieved, which enabled construction of a QC filtered DIA assay library consisting of nearly 4000 proteins that are represented by at least 3 unique peptides, each by at least 4 transitions. This library was used to compare protein abundances of two field populations and one lab-reared B. schlosseri population. Relative protein abundance data indicate that lab-rearing leads to greater proteome changes than those observed between distantly located field populations in Southern California and Northern Washington. STRING analysis revealed functions that are enriched in specific populations and proteins that are least and most variable within a given population.

ORGANISM(S): Botryllus Schlosseri

SUBMITTER: Dietmar Kültz

PROVIDER: PXD043046 | panorama | Thu Feb 29 00:00:00 GMT 2024

REPOSITORIES: PanoramaPublic

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Deep quantitative proteomics of North American Pacific coast star tunicate (Botryllus schlosseri).

Kültz Dietmar D Gardell Alison M AM DeTomaso Anthony A Stoney Greg G Rinkevich Baruch B Rinkevich Yuval Y Qarri Andy A Dong Weizhen W Luu Brenda B Lin Mandy M

Proteomics 20240224 15

Botryllus schlosseri, is a model marine invertebrate for studying immunity, regeneration, and stress-induced evolution. Conditions for validating its predicted proteome were optimized using nanoElute® 2 deep-coverage LCMS, revealing up to 4930 protein groups and 20,984 unique peptides per sample. Spectral libraries were generated and filtered to remove interferences, low-quality transitions, and only retain proteins with >3 unique peptides. The resulting DIA assay library enabled label-free quan ...[more]

PMID: 38400697

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Project description:Data independent acquisition-mass spectrometry (DIA-MS) coupled with liquid chromatography is a promising approach for rapid, automatic sampling of MS/MS data in untargeted metabolomics. However, wide isolation windows in DIA-MS generate MS/MS spectra containing a mixed population of fragment ions together with their precursor ions. This precursor-fragment ion map in a comprehensive MS/MS spectral library is crucial for relative quantification of fragment ions uniquely representative of each precursor ion. However, existing reference libraries are not sufficient for this purpose since the fragmentation patterns of small molecules can vary in different instrument setups. Here we developed a bioinformatics workflow called MetaboDIA to build customized MS/MS spectral libraries using a user's own data dependent acquisition (DDA) data and to perform MS/MS-based quantification with DIA data, thus complementing conventional MS1-based quantification. MetaboDIA also allows users to build a spectral library directly from DIA data in studies of a large sample size. Using a marine algae data set, we show that quantification of fragment ions extracted with a customized MS/MS library can provide as reliable quantitative data as the direct quantification of precursor ions based on MS1 data. To test its applicability in complex samples, we applied MetaboDIA to a clinical serum metabolomics data set, where we built a DDA-based spectral library containing consensus spectra for 1829 compounds. We performed fragment ion quantification using DIA data using this library, yielding sensitive differential expression analysis. </br></br> Serum metabolome of 40 age-related macular degeneration patients and 20 control samples was analyzed using untargeted mass spectrometry. We used data dependent acquisition data to build a MS/MS spectral assay library for more than 1,000 compounds and performed targeted extraction of MS2 ion chromatograms from data independent acquisition analysis.

Dataset Information

Deep quantitative proteomics of North American Pacific coast star tunicate

Publications

Deep quantitative proteomics of North American Pacific coast star tunicate (Botryllus schlosseri).

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