Project Summary
Advances in powerful research technologies such as mass spectrometry have enabled researchers to not only
catalogue the many variants of the protein complement of an organism or organelle system, but more recently,
using quantitative proteomic tools, to specifically quantitate each protein component both spatially and temporally
within the cell. The acquisition of the ThermoFisher Fusion Lumos Orbitrap mass spectrometer (MS) is
crucial to the research infrastructure at the Institute for Systems Biology (ISB) and the collaborative
partnership with the Center for Infectious Disease Research (CIDR). This instrument will enable the
development and broad dissemination of next-generation technology and instrumentation to
researchers ranging from Ph.D. students and post-docs, to experienced PI's at ISB and CIDR and other
collaborating institutes. Used by experimentalists and computational biologists, the data collected with the
proposed instrumentation will enhance the understanding of protein function and action through identification
and quantitation of peptides, their posttranslational modifications and will ultimately inform researchers of
proteome dynamics that are responsible for an organism's form and functions.
The acquisition of the ThermoFisher Fusion Lumos Orbitrap mass spectrometer will substantially improve our
ability to carry out large-scale proteomics experiments at unprecedented depths of proteome coverage that
includes novel PTM analysis such as phosphorylation, ADP-Ribosylation, carbohydrate analysis, and
quantitative proteomics and metabolomics in our research programs. Our request for a Fusion Lumos Orbitrap
MS will enable researchers to perform in-depth studies of the protein networks underlying critical cellular
processes, such as innate immunity, peroxisome function, cell-cell signaling via secreted vesicles and responses
to DNA damage, make inferences from the interacting network of genes and their transcripts important to
understanding the biological context of disease progression. The proposed instrument is critical for the success
of such experiments because of its unique capabilities, including the ability to screen out chemical background
noise, sequence greater numbers of proteotypic and metabolic signatures of target proteins and metabolites per
unit time utilizing very high efficiency chromatography and fast MS scanning rates. We will be able to identify
post-translational modifications with very high specificity and sensitivity not currently achievable with other mass
spectrometers in house or nearby. Currently, ISB and CIDR have access to a previous generation (several
generations older) ThermoFisher LTQ-Velos-Pro Orbitrap Elite MS with ISB staff providing expertise in
experimental design, data acquisition and interpretation however, our needs far exceed our current capabilities.
The planned acquisition and implementation plan described is crucial for the development and completion of
research currently underway at each institute and with their respective collaborating partners.