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IsoTopics™ – July 2016

NeuCode Proteomics and Bap1 Regulation of Metabolism | Cambridge Isotope Laboratories, Inc.

Capitalizing on the merits of neutron encoding (NeuCode), Coon J et al. applied this metabolic-labeling technique to genetically engineered mice models to investigate mammalian biology in vivo. After preliminary experiments on the labeling efficiency and quantitative accuracy, three tissues (namely liver, pancreas, and spleen) were selected for further evaluation of Bap1, a deubiquitinating enzyme with cancer association. This research highlighted the utility of 4-plex NeuCode in mammals and revealed Bap1 to be a key regulator of metabolic homeostasis.

Abstract

We introduce neutron-encoded (NeuCode) amino acid labeling of mice as a strategy for multiplexed proteomic analysis in vivo. Using NeuCode, we characterize an inducible knockout mouse model of Bap1, a tumor suppressor and deubiquitinase whose in vivo roles outside of cancer are not well established. NeuCode proteomics revealed altered metabolic pathways following Bap1 deletion, including profound elevation of cholesterol biosynthetic machinery coincident with reduced expression of gluconeogenic and lipid homeostasis proteins in liver. Bap1 loss increased pancreatitis biomarkers and reduced expression of mitochondrial proteins. These alterations accompany a metabolic remodeling with hypoglycemia, hypercholesterolemia, hepatic lipid loss, and acinar cell degeneration. Liver-specific Bap1 null mice present with fully penetrant perinatal lethality, severe hypoglycemia, and hepatic lipid deficiency. This work reveals Bap1 as a metabolic regulator in liver and pancreas, and it establishes NeuCode as a reliable proteomic method for deciphering in vivo biology. 

Baughman JM, Rose CM, Kolumam G, Webster JD, Wilkerson EM, Merrill AE, Rhoads TW, Noubade R, Katavolos P, Lesch J, Stapleton DS, Rabaglia ME, Schueler KL, Asuncion R, Domeyer M, Zavala-Solorio J, Reich M, DeVoss J, Keller MP, Attie AD, Hebert AS, Westphall MS, Coon JJ, Kirkpatrick DS, Dey A.

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