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Ion Mobility-Assisted Free Radical Initiated Peptide Sequencing

ABSTRACT: Free radical-initiated peptide sequencing (FRIPS), a TEMPO-assisted mass spectrometry, has shown potential as a peptide tandem mass spectrometry (MS/MS) tool that can be an alternative to electron-based mass spectrometry and the traditional collision-based methods. However, a couple of instrumentation barriers need to be overcome before this approach can be a practical tool. One of the barriers is the limitation of the FRIPS-based technique to ion trap mass spectrometry or an orbitrap due to the MS3 workflow. To overcome this challenge, the work introduces a recently developed activation within the trapped ion mobility mass spectrometry (TIMS) device that was found to initiate the radical species. More importantly, the generated product ion was mobility separated, which increased the sequence coverage. Overall, coupling the activation within the TIMS cell with the para-TEMPO-Bz-based FRIPS mass spectrometry offers another practical approach that can be utilized for peptide MS/MS analysis

INSTRUMENT(S): timsTOF

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Nicholas Borotto

PROVIDER: MSV000095935 | MassIVE |

REPOSITORIES: MassIVE

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Ion Mobility-Assisted Free Radical Initiated Peptide Sequencing v2

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Project description:RATIONALE In spatial proteomics, matrix-assisted laser desorption/ionization (MALDI) imaging enables rapid and cost-effective peptide measurement. Yet, in situ peptide identification remains challenging. Therefore, this study aims to integrate the trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) into MALDI imaging of peptides to enable multiplexed MS/MS imaging. METHODS An initial MALDI TIMS MS1 survey measurement is performed, followed by a manual generation of a precursor list containing mass over charge values and ion mobility windows. Inside the dual TIMS system, submitted precursors are trapped, separately eluted by their ion mobility and analyzed in a quadrupole time-of-flight device, thereby enabling multiplexed MALDI MS/MS imaging. Finally, precursors are identified by peptide to spectrum matching. RESULTS This study presents the first multiplexed MALDI TIMS MS/MS imaging (iprm-PASEF) of tryptic peptides. Its applicability is showcased on two histomorphologically distinct tissue specimens in a 4-plex and 5-plex setup. Precursors were successfully identified by the search engine MASCOT in one single MALDI imaging experiment for each respective tissue. Peptide identifications were corroborated by liquid-chromatography tandem mass spectrometry experiments and fragment colocalization analyses. CONCLUSIONS In the present study, we demonstrate the feasibility of TIMS-based MALDI MS/MS imaging for the multiplexed and rapid identification of tryptic peptides in a spatial manner. Hence, it represents a first step towards the integration of MALDI imaging into the emerging field of spatial proteomics. The datasets will be uploaded separately. This data set contains the PDX MSMS data. There are two tissues vissible on the.tif image. The relevant one is on the left side of the slide (away from the barcode).

2024-11-11 | MSV000096376 | MassIVE

Spatial Proteomics by Trapped Ion Mobility supported MALDI MS/MS Imaging: A First Glance into Multiplexed and Spatial Peptide Identification

Project description:RATIONALE In spatial proteomics, matrix-assisted laser desorption/ionization (MALDI) imaging enables rapid and cost-effective peptide measurement. Yet, in situ peptide identification remains challenging. Therefore, this study aims to integrate the trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) into MALDI imaging of peptides to enable multiplexed MS/MS imaging. METHODS An initial MALDI TIMS MS1 survey measurement is performed, followed by a manual generation of a precursor list containing mass over charge values and ion mobility windows. Inside the dual TIMS system, submitted precursors are trapped, separately eluted by their ion mobility and analyzed in a quadrupole time-of-flight device, thereby enabling multiplexed MALDI MS/MS imaging. Finally, precursors are identified by peptide to spectrum matching. RESULTS This study presents the first multiplexed MALDI TIMS MS/MS imaging (iprm-PASEF) of tryptic peptides. Its applicability is showcased on two histomorphologically distinct tissue specimens in a 4-plex and 5-plex setup. Precursors were successfully identified by the search engine MASCOT in one single MALDI imaging experiment for each respective tissue. Peptide identifications were corroborated by liquid-chromatography tandem mass spectrometry experiments and fragment colocalization analyses. CONCLUSIONS In the present study, we demonstrate the feasibility of TIMS-based MALDI MS/MS imaging for the multiplexed and rapid identification of tryptic peptides in a spatial manner. Hence, it represents a first step towards the integration of MALDI imaging into the emerging field of spatial proteomics. The datasets will be uploaded separately. This data set contains the Kidney MSMS data. There is an additional Liver tissue vissible on the .tif image. This was used for another experiment

2024-11-11 | MSV000096375 | MassIVE

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