A Proof of Concept to Bridge the Gap between Mass Spectrometry Imaging, Protein Identification and Relative Quantitation: MSI~LC-MS/MS-LF
AbstractMass spectrometry imaging (MSI) is a powerful tool to visualize the spatial distribution of molecules on a tissue section. The main limitation of MALDI-MSI of proteins is the lack of direct identification. Therefore, this study focuses on a MSI~LC-MS/MS-LF workflow to link the results from MALDI-MSI with potential peak identification and label-free quantitation, using only one tissue section. At first, we studied the impact of matrix deposition and laser ablation on protein extraction from the tissue section. Then, we did a back-correlation of the m/z of the proteins detected by MALDI-MSI to those identified by label-free quantitation. This allowed us to compare the label-free quantitation of proteins obtained in LC-MS/MS with the peak intensities observed in MALDI-MSI. We managed to link identification to nine peaks observed by MALDI-MSI. The results showed that the MSI~LC-MS/MS-LF workflow (i) allowed us to study a representative muscle proteome compared to a classical bottom-up workflow; and (ii) was sparsely impacted by matrix deposition and laser ablation. This workflow, performed as a proof-of-concept, suggests that a single tissue section can be used to perform MALDI-MSI and protein extraction, identification, and relative quantitation. View Full-Text
- Supplementary File 1:
Supplementary (XLSX, 516 KB)
Externally hosted supplementary file 1
Description: Supplementary Materials: Supplementary Data 1: Table of identified proteins in the 8 individuals within the three conditions: ‘On tissue’, ‘SA-coated’, and ‘Post-MSI’ extractions, and their main cellular compartment. Supplementary Data 2: Classification of proteins according to their main and intermediate cellular compartment. Supplementary Data 3: Comparison of proteins identified by ‘Muscle homogenate’ proteomic workflow. Supplementary Data 4: Label-free quantitation analysis performed on proteins extracted in situ ‘Post-MSI’ and MALDI-MSI intensities.
Scifeed alert for new publicationsNever miss any articles matching your research from any publisher
- Get alerts for new papers matching your research
- Find out the new papers from selected authors
- Updated daily for 49'000+ journals and 6000+ publishers
- Define your Scifeed now
Théron, L.; Centeno, D.; Coudy-Gandilhon, C.; Pujos-Guillot, E.; Astruc, T.; Rémond, D.; Barthelemy, J.-C.; Roche, F.; Feasson, L.; Hébraud, M.; Béchet, D.; Chambon, C. A Proof of Concept to Bridge the Gap between Mass Spectrometry Imaging, Protein Identification and Relative Quantitation: MSI~LC-MS/MS-LF. Proteomes 2016, 4, 32.
Théron L, Centeno D, Coudy-Gandilhon C, Pujos-Guillot E, Astruc T, Rémond D, Barthelemy J-C, Roche F, Feasson L, Hébraud M, Béchet D, Chambon C. A Proof of Concept to Bridge the Gap between Mass Spectrometry Imaging, Protein Identification and Relative Quantitation: MSI~LC-MS/MS-LF. Proteomes. 2016; 4(4):32.Chicago/Turabian Style
Théron, Laëtitia; Centeno, Delphine; Coudy-Gandilhon, Cécile; Pujos-Guillot, Estelle; Astruc, Thierry; Rémond, Didier; Barthelemy, Jean-Claude; Roche, Frédéric; Feasson, Léonard; Hébraud, Michel; Béchet, Daniel; Chambon, Christophe. 2016. "A Proof of Concept to Bridge the Gap between Mass Spectrometry Imaging, Protein Identification and Relative Quantitation: MSI~LC-MS/MS-LF." Proteomes 4, no. 4: 32.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.