Baughman Joshua M, Rose Christopher M, Kolumam Ganesh, Webster Joshua D, Wilkerson Emily M, Merrill Anna E, Rhoads Timothy W, Noubade Rajkumar, Katavolos Paula, Lesch Justin, Stapleton Donald S, Rabaglia Mary E, Schueler Kathy L, Asuncion Raymond, Domeyer Melanie, Zavala-Solorio Jose, Reich Michael, DeVoss Jason, Keller Mark P, Attie Alan D, Hebert Alexander S, Westphall Michael S, Coon Joshua J, Kirkpatrick Donald S, Dey Anwesha
Department of Protein Chemistry, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
Cell Rep. 2016 Jul 12;16(2):583-595. doi: 10.1016/j.celrep.2016.05.096. Epub 2016 Jun 30.
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.
我们引入了小鼠的中子编码（NeuCode）氨基酸标记法，作为体内多重蛋白质组分析的一种策略。利用NeuCode，我们对Bap1的诱导型基因敲除小鼠模型进行了表征，Bap1是一种肿瘤抑制因子和去泛素化酶，其在癌症之外的体内作用尚未完全明确。NeuCode蛋白质组学揭示了Bap1缺失后代谢途径的改变，包括胆固醇生物合成机制的显著升高，同时肝脏中糖异生和脂质稳态蛋白的表达降低。Bap1缺失增加了胰腺炎生物标志物，并降低了线粒体蛋白的表达。这些改变伴随着代谢重塑，出现低血糖、高胆固醇血症、肝脏脂质流失和腺泡细胞变性。肝脏特异性Bap1基因敲除小鼠表现出完全显性的围产期致死率、严重低血糖和肝脏脂质缺乏。这项工作揭示了Bap1作为肝脏和胰腺中的代谢调节因子，并将NeuCode确立为一种可靠的蛋白质组学方法，用于解读体内生物学。
