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Integrated mass spectrometry strategies to decipher metabolite-protein cross regulation

Award Number: R35GM162269
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 03/02/2026
PERIOD OF PERFORMANCE END DATE: 12/31/2030

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Integrated mass spectrometry strategies to decipher metabolite-protein cross regulation - Integrated mass spectrometry strategies to decipher protein-metabolite cross regulation Cellular metabolism relies on protein enzymes to convert substrates to products, working in interconnected networks to generate important biological precursors, remove toxic waste, balance redox and defend energy potential. However, the study of metabolic regulation in both healthy and disease states has proven challenging in part due to the intricate cross-regulation between metabolites and proteins. Current strategies to characterize metabolic regulation have generally focused on characterizing metabolites or protein enzymes, but not both. Consequently, how metabolites and proteins interact in the cellular context remains an open question in the field of metabolism and cellular biology. To bridge this gap, the Skinner lab is focused on characterizing the interactions between metabolites and proteins in the cellular context by using mass spectrometry to integrate existing multi-tiered methods in addition to developing new techniques to enhance the characterization and scope of both protein and metabolite measurements. Our primary research focus is on the regulation of proteins and metabolism by redox cofactors through signaling of sulfur-containing metabolites (Area 1) and the effect of vitamin B6 on metabolic and redox regulation as it is converted to protein cofactor pyridoxal-5-phosphate (PLP) (Area 2). These two areas each represent critical intersections of metabolome and proteome that play a key role across a variety of disease states. To better understand these two ranges of metabolite and protein cross-regulation, we will apply top-down proteomics and native mass spectrometry analyses of intact protein complexes coupled with full-scan and stable isotope labeling metabolomics to simultaneously probe metabolites and proteins. In Area 1, we will characterize how the abundance and redox state of specific thiol metabolites and cellular nicotinamide adenine dinucleotide phosphate (NADP+) redox state can modulate glycolysis and other core metabolic pathways Protein thiols have been demonstrated to be covalently modified at high stoichiometry with a variety of different groups, making this a likely axis for metabolic regulation. In Area 2, we will examine another key intersection between metabolite and protein regulation; the metabolism of vitamin B6 to its active form PLP prior to acting as a cofactor for 53 human enzymes. Specifically, we will characterize how some PLP-containing enzymes seem more resistant to extended B6 deficiency than others and how changes in subcellular nicotinamide adenine dinucleotide (NAD+) redox state can affect PLP levels and vice versa. This project integrates metabolomics with native mass spectrometry and proteomics and applies it to two pressing questions in the field of systems biology and regulation that lie at the intersection of metabolome and proteome.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $434,000 )
2026	2026	NORTHEASTERN UNIVERSITY	360 HUNTINGTON AVE	BOSTON	MA	02115	SUFFOLK	USA	Biomedical Research and Research Training	000	1	3/2/2026	NEW	$434,000
														Subtotal = $434,000

Grand Total All Awards = $434,000

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Integrated mass spectrometry strategies to decipher metabolite-protein cross regulation

Award Number: R35GM162269

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 03/02/2026

PERIOD OF PERFORMANCE END DATE: 12/31/2030

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