The secret life of plants: intact protein mass spectrometry imaging by nanospray desorption electrospray ionization (nanoDESI)
EMSL Project ID
60273
Abstract
Intact protein mass spectrometry imaging (MSI) is a niche field of study which combines the specificity of mass spectrometry (MS) with the capabilities of visualization as in microscopy. Proteomics applications within MSI at large have been focused upon the development and application to clinical mammalian research, principally via matrix-assisted laser desorption/ ionization (MALDI). While MALDI has proven an effective tool for protein and proteoform mapping; high endogenous water content, hard cell walls, and large air gaps in plant tissues cause issues for plant tissue stability under requisite vacuum conditions for MALDI-MSI. For these reasons, ambient protein MSI a highly desirable alternative. EMSL has previously pioneered the development of nanospray desorption electrospray ionization (nanoDESI), a MSI technique which is capable of mapping of biomolecules under ambient conditions. Several literature reports have since utilized nanoDESI-MSI for the mapping of protein and proteoform distributions within murine tissues, however, no studies have completed an evaluation of nanoDESI on plant samples. The proposed research will couple nanoDESI to various mass spectrometers within the Fourier transform mass spectrometry (FT-MS) suite at EMSL for a formative evaluation of intact protein nanoDESI-MSI on soybean root nodules. This study will include a validation of methodologies on mammalian tissue, leading to the MSI of soybean root nodules which host a highly dynamic endosymbiotic relationship with rhizobia bacteria that are responsible for the fixation of nitrogen. The preservation of cellular spatial distributions and colocalizations of proteins via imaging under ambient conditions will lend itself to a highly complementary approach to bottom-up and top-down proteomics. Further developing a knowledge base which will later be applied at PNNL to other plant, bacterial, and fungal samples. This LDRD funded research through QuickStarter will have broad ranging implications.
Project Details
Start Date
2021-12-15
End Date
2022-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
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